JP2772372B2 - Cyclosporins and their pharmaceutical uses - Google Patents

Abstract

Cyclosporins wherein the residue at the 1-position (typically -MeBmt- or -dihydro-MeBmt-) is 3 min O-acylated or 3 min -oxo or -C1-4alkoxyimino substituted, or wherein the residue at the 2-position is beta -O-acyl or beta -oxo substituted, or wherein the residue at the 2-position is -Ile-, or wherein the residue at the 11-position is -MeAla-, -MeIle- or -MealloIle- as well as various naturally occurring cyclosporins/dihydro-derivatives thereof, are useful in reversing resistance to chemotherapy, in particular resistance to cytostatic or anti-neoplastic therapy. Various of these cyclosporins and intermediates for their production are novel. Intermediates wherein the residue (e.g. -MeBmt-, -dihydro-MeBmt- etc.) at the 1-position is 8 min -alkoxy of 7 min -desmethyl-7 min -hydrocarbyl substituted are novel and useful as immunosuppressants, anti-inflammatory and anti-parasitic agents.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention relates to a novel use of cyclosporins, in particular, a novel pharmaceutical use and a novel cyclosporin which is a novel compound per se.

[0002]

BACKGROUND OF THE INVENTION Cyclosporins, to varying degrees, generally have pharmacological, in particular immunosuppressive, anti-inflammatory and / or antiparasitic activity, and cyclic poly-N-types having a unique structure. Includes types of methylated undecapeptides. The first isolated cyclosporins, also known as "cyclosporin A", are currently marketed under the trademark SANDIMMUN, a natural fungal metabolite "cyclosporin" (Ciclosporin or Cyclosporine).
Met. “Cyclosporin” has the formula (A)

Embedded image [In the formula, -MeBmt- is the formula (B)

Embedded image (Wherein -xy- is trans -CH = CH-,
The 2 ', 3' and 4 'positions have the configurations S, R and R, respectively), N-methyl- (4R) -4-buta-2
E-en-1-yl-4-methyl- (L) threonyl residue].

Since the first discovery of "cyclosporine", a wide variety of natural cyclosporins have been isolated and identified,
Many more unnatural cyclosporins have been produced by the application of complete or semi-synthetic means or modified culture techniques. That is, currently, cyclosporins include many types, for example, natural cyclosporins AZ [reference, traversal, etc., 1, "Helvetica simica acta" (Helv. Chim. A
cta), 60, pp. 1247-1255 (1977), Traver et al., 2, `` Helvetica Simica Acta '' (Helv.
Chim. Acta), 65, 1655-1667 (1982)
), Kobel et al., `` European Journal of
Applied Microbiology and Biotechnology ”(Europ. J. Applied Microbiology and Biotechnology)
technology), 14, 273-240 (1982)
And von Baltburg, "Progress in
Progress in Allergy, Volume 38, 28-4
5 (1986)] and various unnatural cyclosporin derivatives and dihydro- and iso-cyclosporins
[The above formula to saturate the -x-y- portion of -MeBmt- residue in (B) -x-y - = - CH 2 -CH 2 - to / above formula
The residue at position 11 of the cyclosporin molecule in (A) and -M
Artificial or synthetic cyclosporins, derivatized cyclosporins (e.g., 3'- of the MeBmt-residue), wherein the linkage of the eBmt-residue is through the 3'-O-atom rather than the α-N-atom.
Acylating the O-atom or introducing another substituent at the α-carbon atom of the sarkosyl residue at position 3), cyclosporins in which the -MeBmt- residue is present in isomeric form (eg -MeB
The configuration across the 6 'and 7' positions of the mt-residue is cis rather than trans) and, for example, using the fully synthetic method of producing cyclosporins developed by Wenger et al. Cyclosporins to be introduced into specific positions of the compounds [for example, Traver et al. 1, Traver et al. 2, and Kobel et al., Supra, US Pat. Nos. 4,108,985, 4,210,581, and 42.
No. 20641, No. 4288431, No. 45543
51 and 4,396,542, EP-A-0034567 and EP-A-0056678, International Patent Publication No. WO 86/02080, Benger 1,
"Transpl. Proc.", Volume 15,
Addendum 1: 2230 (1983), Bengar 2, "Angevante Chemie, International Edition.
In English "(Angew. Chem. Int. Ed.), 24
Vol. 77, 1985, and Benger 3, "Progress in the Chemistry of Organic Products"
Organic Natural Products), 50 volumes, 123 pages (19
1986)].

[0004] That is, the types of cyclosporins presently are very large, for example, [Thr] ^2- , [Val] ^2- , and [Nva].
^2- and [Nva] ^2- [Nva] ⁵ -cyclosporin (also known as cyclosporins C, D, G and M, respectively), [3-O-acetyl-MeBmt] ¹ -cyclosporin
(Also known as cyclosporin A acetate), [dihydro-MeBmt] ^1- [Val] ² -cyclosporin
(Also known as dihydro-cyclosporin D),
[Iso-MeBmt] ^1- [Nva] ² -cyclosporin (also known as isocyclosporin G), [(D) Ser] ⁸ -cyclosporin, [MeIle] ¹¹ -cyclosporin, [(D) M
eVal] ¹¹ -cyclosporin (also known as cyclosporin H), [MeAla] ⁶ -cyclosporin, [(D) P
ro] ³ -cyclosporin and the like.

[These definitions are made according to the conventional nomenclature of cyclosporins by indicating the structure of "cyclosporin" (ie cyclosporin A) throughout this specification (including the claims). This first shows the amino acid residues present that are different from those present in “cyclosporin” (eg, [(D) Pro] ³ indicates that the cyclosporin in question is at position 3 with a-( D) indicating that it has Pro-) and then applying the term "cyclosporin" to characterize the remaining residues as being present in "cyclosporin".

The first residue of "cyclosporin"-MeBmt
-Was unknown before the discovery of cyclosporine.
That is, this residue and its variants or modifications
(Eg, below) are generally specific to cyclosporins. In general, variants or alternatives to [MeBmt] ¹ are defined by indicating a -MeBmt- structure. That, -x-y- portion [the formula (B) Reference is -CH ₂ -CH ₂ - For dihydro cyclosporins has been reduced to the 1-position residues are "- dihydro -MeB
mt- ". If the configuration across the -xy- moiety is cis rather than trans, then this residue is a "-
Cis-MeBmt- ".

When deleting the -MeBmt- residue,
This is because the determinants "-MeBmt-", "-dihydro-MeBmt-
"-", "Cis-MeBmt-", etc. are indicated by defining the position of the deletion, indicating the deletion using the modifier "des", and then defining the group or atom to be deleted. That is, "-
N-desmethyl-MeBmt- ","-3'-desoxy-M
eBmt- "and"3'-desoxy-4'-desmethyl -MeBmt- "are each formula (B ^1), a residue represented by (B ²⁾ and (B ^3).

[0008]

Embedded image

For example, when substituting a position or group for -MeBmt-, this is represented in a conventional manner by defining the position and nature of the substitution. That,-3'-O-acetyl -MeBmt- the formula (B) [However, 3'-OH group is acetylated _{(3'-O-COCH 3)} ] is a residue represented by. For example, when substituting a substituent or group of -MeBmt-, this is done by i) indicating the position of the group substituted by the above "death-term" and ii) defining the substituent. That is, -7'-desmethyl-7'-phenyl-MeBmt- is represented by the formula (B) [where the terminal (8 ')
A methyl group is substituted by phenyl]. 3'-desoxy-3'-oxo-MeBmt
-Is a residue represented by the above formula (B) (provided that the 3'-OH group is substituted by = O).

Furthermore, abbreviations such as -Ala-, -Me
The amino acid residues represented by Val-, -αAbu-, etc.
In accordance with convention, unless otherwise specified, as in the case of "-(D) Ala-", for example, it is defined to have the (L) -configuration.
Residue abbreviations preceded by "Me" as in "-MeLeu-" represent α-N-methylated residues. In the art, at each residue of the cyclosporin molecule, the residue at position 1 -Me
Starting from Bmt-, -dihydro-MeBmt-, etc., numbers are given in clockwise direction. This specification (including the claims) consistently uses the same sequence of numbers. ]

[0011]

SUMMARY OF THE INVENTION Cyclosporins are
Due to their unique medicinal properties, they have received a great deal of attention not only from the medical and university community, but also from non-professional press. Currently, cyclosporine itself is routinely used in preventing rejection after allogeneic organ (eg, heart, cardiopulmonary, kidney, and bone marrow) transplantation, and more recently in the treatment of various autoimmune and related diseases and conditions. Extensive work has also been undertaken to study potential uses in treating various parasitic diseases and infectious diseases such as coccidioidomycosis, malaria and schistosomiasis. Current,
Numerous reports of work on the potential use of numerous other known cyclosporins in these or related indications have been described in the literature.

According to the present invention, now surprisingly,
Certain cyclosporins have been found to be useful in enhancing sensitivity to other chemotherapies, and in particular, in reversing resistance to other chemotherapies, whether inducible or intrinsic.

[0013] The high resistance to chemotherapy after short or long term treatment, generally after administration of relatively long chemotherapeutic drugs, is a widespread phenomenon that has been recognized for quite some time and is widely recognized in the art. It is stated in the document. Classic examples include enhanced or induced resistance of parasites, especially bacterial, viral or protozoan microorganisms, after prolonged or extensive drug therapy with individual drug substances. In such cases,
Infectious microorganisms become more or less resistant to drug substances over time, with concomitant difficulties in fighting or treating. Similar phenomena are observed with chemotherapeutic treatment of cancer, for example, treatment of carcinoma, sarcoma or other tumors or malignant tumors.

[0014] As a means of reducing, inhibiting or limiting the growth or metastasis of tumors, for example, the administration of drug substances, in particular anti-neoplastic or cytotoxic drugs, such as colchicine, etoposide, tenoposide, adriamycin, daunorubicin and vincristine, the treatment of cancer. Chemotherapy treatment
It remains the most important study of the treatment of various types of cancer. However, it has generally been found that tumors may initially be sensitive to treatment, but as treatment continues, resistance to such treatment develops, resulting in a diminished therapeutic effect. As a general rule, multi-drug resistance usually follows, especially when pleiotropic or multi-drug therapy is employed, especially in the treatment of late-stage or end-stage cancer.
For example, if a certain form of chemotherapy treatment is performed for a long time,
Similar disorders arise, for example, resulting in resistant strains of microorganisms that have developed intrinsic or endogenous resistance. Again, one often encounters certain forms of cancer or tumor that exhibit endogenous resistance or low levels of sensitivity to conventional anti-neoplastic or cytotoxic drug therapies.

[0015]

According to the present invention, according to the present invention, the cyclosporins described below are useful for increasing the sensitivity to chemotherapeutic drug therapy or enhancing its effects, and in particular, for various types (eg, (Innate or congenital) chemotherapy drug resistance, or has been found to be useful for enhancing or restoring sensitivity to applied drug therapy. Forms of chemotherapeutic drug therapy to which this invention can be applied include, for example, antiparasitic, such as antiviral, antibacterial or antigenic animal chemotherapy, and especially anti-neoplastic or cytostatic chemotherapy.

[0016] Cyclosporins suitable for use in the present invention can be defined under the various classes shown below. Class 1 A cyclosporin in which the 3'-carbon atom of the 1-position residue or the β-carbon atom of the 2-position residue is O-acyl or oxo substituted. Preferred cyclosporins of this class are: (1a) When the 3′-carbon atom of the residue at position 1 is O-acyl substituted. If the 3′-carbon atom of the residue at position 1 is O-acyl substituted, the residue at position 2 may also be β-O-acylated. Preferred groups of (1a) type cyclosporins are as follows: (1a ¹ ) wherein the residue at the 1-position is -3′-O-acyl-MeBmt- or -3′-acyl-dihydro-MeBm represented by the formula (I)
If it is a t-residue.

Embedded image (Wherein -xy- is -CH ₂ CH ₂ -or trans-C
H = CH-, ACYL ¹ represents an acyl group)

[0017] Suitable acyl groups as ACYL ¹ in formula (I) are those wherein R ₁ -CO- and R ₂ -O-CO- (wherein, R ₁
The C ₁ - ₄ alkyl or C ₁ - ₄ azidoalkyl and R _2
4 groups represented by alkyl as), for example, acetyl, - C ₁ is
4-Azidobutanoyl or methoxycarbonyl. Preferably ACYL ¹ is a group represented by the formula R ₁ -CO-, in this case R ₁ is preferably C ₁ - ₄ alkyl. Most preferably, R ₁ —CO— is acetyl.

The alkyl groups which are or contain R ₁ and R ₂ can be branched or straight-chain. Preferably they are linear. R ₁ is most preferably methyl.

Particularly preferred cyclosporins of the formula (1a ¹ ) are those of the formula (II) or (II ′).

Embedded image [Wherein A is the residue defined in the above (1a ¹ ), and B is -α
Abu-, -Thr-, -Val-, -Nva- or? -O-
X is -Sar- or a residue of an acyl-α-amino acid;
(D)-an optically active α-N-methylated α amino acid residue having a configuration, Y is -Val- or, furthermore, B is-
When Nva-, -Nva- and W are β-hydroxy- or β-O-acyl- having the (D) -configuration.
B is a β-O-acyl-α-
When it is an amino acid residue, it generally has the (L) configuration. Preferably it is a residue of formula (III).

Embedded image In the formula, ACYL ² represents an acyl group. Preferably it is an O-acyl- (L) -threonyl residue. W is preferably-(D) Ser- or-(D) Thr- or a compound of formula (III ')
And O-acyl- (D) Ser- or O-acyl- (D) Thr-.

Embedded image In the formula, ACYL ² has the same meaning as in formula (III), and R is hydrogen or methyl.

[0020] Preferred acyl groups as ACYL ² in formula (III) and (III ') is, (wherein, R ₃ is C ₁ - ₄ alkyl) wherein R ₃ -CO- a group represented by. The alkyl group as R ₃ can be branched or straight. Preferably they are linear. Suitably ACYL ² is acetyl.

When X in formula (II) is other than -Sar-, it is preferably-(D) Ala-.

Examples of this class of cyclosporins suitable for use in accordance with the present invention are as follows: 1.1 [3′-O-acetyl-MeBmt] ¹ -cyclosporin, 1.2 [3′-O-acetyl-MeBmt] ^1- [Val] ² -cyclosporin, 1.3 [3′-O-acetyl-MeBmt] ^1- [Thr ] ² - Ciclosporin, 1.4 [3'-O- acetyl-MeBmt] ¹ - [Nva] ² - Ciclosporin, 1.5 [3'-O- acetyl ^{-MeBmt] 1 - [Nva] 2} - [Nva]
⁵ -cyclosporin, 1.6 [3'-O-acetyl-MeBmt] ¹ -[(D) Ala] ³ -cyclosporin, 1.7 [3'-O-acetyl-MeBmt] ^1- [Nva] ² -[(D)
Ala] ³ -cyclosporin, 1.8 [3'-O-acetyl-MeBmt] ¹ -[(D) MeVal] ¹¹
-Cyclosporin, 1.9 [3'-O-acetyl-MeBmt] ^1- [Val] ¹¹ -cyclosporin, 1.10 [3'-O-acetyl-dihydro-MeBmt] ¹ -cyclosporin, 1.11 [3'-O-methoxycarbonyl- MeBmt] ¹ -cyclosporin, 1.12 [3′-O- (4-azidobutanoyl) -MeBmt] ¹ −
Cyclosporin, 1.13 [3'-O-acetyl-MeBmt] ^1- [O-acetyl-Thr] ² -cyclosporin, 1.14 [3'-O-acetyl-N-desmethyl-MeBmt] ¹
-[O-acetyl-Thr] ² -cyclosporin, and 1.15 [3'-O-acetyl-MeBmt] ^1- [O-acetyl- (D) Ser] ⁸ -cyclosporin.

Cyclosporins suitable for use according to the present invention include all of the above cyclosporins 1.1-1.10 and
1.15, and especially cyclosporins 1.1, 1.2, 1.3, 1.
4, 1.10 and 1.15. Another group of (1a) type cyclosporins are as follows: (1a ²⁾ formula,-3'-O-acyl--8'-C ₁ to position 1 residue represented by the formula (IV) - ₈
Alkoxy-cis-MeBmt- or -dihydro-
If MeBmt-residue.

Embedded image (Wherein, -x-y- is cis -CH = CH- or -CH ₂ -
CH ₂ -, R ₄ is C ₂ - ₉ alkoxymethyl and ACYL ¹
The Particularly suitable are acyl groups in acyl groups) Formula (IV), wherein R ₅ -CO- (wherein, R ₅ is C ₁ - is a group represented by a _4-alkyl).

The alkyl group as R ₅ and the alkyl moiety containing R ₄ can be branched or straight chain. Preferably they are linear. Preferably R ₄ is C ₂ - is a ₅ alkoxymethyl. ACYL ¹ in formula (IV) is preferably acetyl.

Particularly preferred cyclosporins of the group (1a ² ) are those of the above formula (II) wherein A is a residue of the above formula (IV),
B has the same meaning as in formula (II), X is -Sar- and Y is -Val-).

Examples of (1a ² ) group cyclosporins suitable for use according to the invention are as follows: 1.16 [3'-O-acetyl-8'-methoxy-cis-Me
Bmt] ¹ -cyclosporin, 1.17 [3′-O-acetyl-8′-t-butoxy-cis-
MeBmt] ¹ -cyclosporin, 1.18 [3′-O-acetyl-8′-methoxy-cis-Me
Bmt] ^1- [O-acetyl-Thr] ² -cyclosporin, 1.19 [3′-O-acetyl-8′-t-butoxy-cis-
MeBmt] ^1- [O-acetyl-Thr] ² -cyclosporin, 1.20 [3′-O-acetyl-8′-methoxy-cis-Me
Bmt] ^1- [Val] ² -cyclosporin and 1.21 [3′-O-acetyl-8′-methoxy-cis-Me
Bmt] ^1- [Nva] ² -cyclosporin.

1a ³ ) wherein the residue at the 1-position is -3'-O-acyl-cis-MeBmt-, -3'-O-acyl-7'-desmethyl-7'-hydrocarbyl--MeBmt- or Cis-MeBmt-residue, where the hydrocarbyl (hydrocarbon residue) moiety contains at least 2 carbon atoms, or -3'-O-acyl-7'-desmethyl-7'-hydrocarbyl-dihydro- When it is a MeBmt residue (provided that the hydrocarbyl moiety contains at least 2 carbon atoms and that any aliphatic groups or moieties present in said hydrocarbyl moiety are saturated).

These residues can be represented by formula (V).

Embedded image (Wherein -xy- is cis or trans -CH = CH-
Or -CH ₂ -CH ₂ - and and, R ₆ is hydrocarbyl, although ACYL ¹ represents an acyl group, provided that, -x-
y- is trans -CH = CH- or -CH ₂ -CH ₂ -
R ₆ contains at least 2 carbon atoms (ie, other than methyl), and -x-y- is -CH ₂ -CH
₂ -, then the aliphatic group or moiety present in a or R ₆ as the R ₆ are assumed to be all saturated)

[0029] Formula (V) Particularly suitable as acyl groups in the formula -R ₅ -CO- (provided that, R ₅ is C ₁ - ₄ alkyl as) a group represented by, in particular acetyl.

The hydrocarbon residue as R ₆ includes aromatic, aliphatic and araliphatic groups, provided that the aliphatic groups and moieties can be branched or straight-chain. Also, such groups may further include substituents such as halogen or hydroxy, or may be unsubstituted.

Suitable hydrocarbon residues for R ₆ are alkyl, alkenyl, alkynyl, phenyl, phenylalkyl, phenylalkenyl and phenylalkynyl,
In particular alkyl, alkenyl and alkynyl groups having up to 20, preferably up to 8 carbon atoms, and phenyl, phenylalkyl and phenylalkynyl groups containing up to 12 carbon atoms.

[0032] Particularly preferred groups R ₆ are phenyl, phenyl - (C ₁ - ₄ alkyl), C ₁ - ₁₀ alkyl, C ₂ - ₁₀ alkenyl and C ₂ - ₁₀ alkynyl, in particular phenyl and C ₁ -
₅ alkyl.

When -xy- in the formula (V) is -CH ₂ -CH _2- , the hydrocarbon residue as R ₆ is such that the aromatic group, the saturated aliphatic group and the aliphatic moiety are saturated. And includes, for example, all of the aforementioned groups belonging to this type described above.

Particularly preferred cyclosporins of this group are those of the above formula (II) wherein A is a residue of the above formula (V) and B is a compound of the formula (V)
X is -Sar- and Y is -Val-
).

Examples of this group of cyclosporins suitable for use according to the invention are as follows: 1.22 [3′-O-acetyl-7′-desmethyl-7′-phenyl-MeBmt] ¹ -cyclosporin, 1.23 [3′-O-acetyl-cis-MeBmt] ¹ -cyclosporin, 1.24 [3′-O-acetyl] -7'-desmethyl-7'-vinyl-cis-MeBmt] ¹ -cyclosporin, 1.25 [3 '
-O-acetyl-7'-desmethyl-7'-vinyl-cis-MeBmt] ^1- [O-acetyl-Thr] ² -cyclosporin, 1.26 [3'-O-acetyl-7'-desmethyl-7'-i −
Pentyl-cis-MeBmt] ¹ -cyclosporin, 1.27 [3′-O-acetyl-7′-desmethyl-7′-phenyl-cis-MeBmt] ¹ -cyclosporin, 1.28 [3′-O-acetyl-7′-desmethyl] -7'-n-
Propyl-cis-MeBmt] ¹ -cyclosporin, 1.29 [3′-O-acetyl-7′-desmethyl-7 ′-(β
-Allyl) -cis-MeBmt] ¹ -cyclosporin, 1.30 [3′-O-acetyl-7′-desmethyl-7′-phenyl-MeBmt] ^1- [Val] ² -cyclosporin, 1.31 [3′-O-acetyl -7'-desmethyl-7'-phenyl-cis-MeBmt] ^1- [Val] ² -cyclosporin, 1.32 [3'-O-acetyl-7'-desmethyl-7'-vinyl-cis-MeBmt] ^1- [ Val] ² -cyclosporin, 1.33 [3'-O-acetyl-7'-desmethyl-7'-vinyl-cis-MeBmt] ^1- [Nva] ² -cyclosporin, and 1.34 [3'-O-acetyl-7 ' -Desmethyl-7 '-(3
-Bromo-n-propyl) -cis-MeBmt] ¹ -cyclosporin.

Another group of class 1 cyclosporins are as follows: (1b) In the formula, when the β-carbon atom of the residue at position 2 is O-acyl substituted, that is, the residue at position 2 is a β-O-acyl-α-amino acid residue, Is the above (1
a) Other than those described in paragraph a).

When the residue at position 2 is a β-O-acyl-α-amino acid residue, this residue generally has the (L) configuration. Preferably, it is a residue of the formula (III ').

Embedded image Wherein ACYL ³ represents an acyl group. Preferably it is an O-acyl- (L) -threonyl residue.

A group suitable as ACYL ³ in the formula (III ") is a group represented by the formula R ₃ --CO-- or R ₇ --CO--R ₈ --CO-- (where R ₃ has the same meaning as in the formula (III), R ₇ is β-position 2
Oxy - (L) -α- residues of cyclosporins through the β- oxygen atom is bonded to the -CO-R ₈ -CO- portion of the amino acid residue is the residue, and R ₈ is C ₁ - ₈ Which is an alkylene).

The alkylene group as R ₈ can be branched or straight-chain. Preferably they are linear. R ₈
Preferably C ₁ - a ₄ alkylene. Acyl groups of the formula R ₃ —CO— are generally preferred, with preferred meanings of R ₃ —CO— being as described above for formula (III).

Particular cyclosporins of this group are those of the formula (II) wherein A is -MeBmt- or -dihydro-M
eBmt- and B are as described in the above item (1b), for example, those represented by the above formula (III '), X is -Sar- and Y is -Val-.

Examples of said cyclosporins suitable for use according to the invention are as follows: 1.35 [O-acetyl-Thr] ² -cyclosporin, and 1.36 1,2-ethanedicarboxylic acid [O-threonyl] ^2-
Cyclosporin diester.

Cyclosporine 1.36 is represented by the formula (VI).

Embedded image Further preferred groups of class 1 cyclosporins are as follows: (1c) In the formula, the 3′-carbon atom of the residue at position 1 is oxo-substituted. If the 3'-carbon atom of the residue at position 1 is oxo substituted, this is preferably of the formula (V
-3'-desoxy-3'-oxo-MeB represented by II)
mt-.

Embedded image Particularly preferred cyclosporins of this group are those of the above formula (II)
(Where A is the residue of the above formula (VII), B is the meaning given by the formula (II), X is -Sar- and Y is -Val-).

Yet another group of class 1 cyclosporins is as follows: (1d) In the formula, when the β-carbon atom of the residue at position 2 is β-oxo substituted, that is, when the residue at position 2 is a β-oxo-α-amino acid residue.

When the residue at position 2 is a β-oxo-α-amino acid residue, this residue generally has the (L) configuration. Preferably it is -α-methylketo-Gly- of formula (VIII).

Embedded image

(1c) suitable for use according to the invention and
Examples of cyclosporins belonging to the group (1d) are as follows. 1.37 [3′-desoxy-3′-oxo-MeBmt] ¹ -cyclosporin, 1.38 [3′-desoxy-3′-oxo-MeBmt] ^1- [V
al] ² -cyclosporin, 1.39 [3'-desoxy-3'-oxo-MeBmt] ^1- [N
va] ² -cyclosporin, 1.40 [α-methylketo-Gly] ² -cyclosporin, and 1.41 [dihydro-MeBmt] ^1- [α-methylketo-Gly] ²
-Cyclosporin. Cyclosporins 1.37-1.39 are particularly preferred.

[0046] Class 2 i) position 1 residue 3'carbon atom C ₁ - ₄ alkoxyimino cyclosporin substituted, for example, position 1 residue -
3'-desoxy -3 '- (C ₁ - ₄ alkoxyimino) -Me
Bmt-residue cyclosporin, or ii) residue at position 2 is (L) -isoleucyl residue, or iii) residue at position 11 is N-methyl- (L) -alanyl, N-
Methyl- (L) -isoleucyl or N-methyl- (L)-
Cyclosporine, which is an alloisoleucyl or N-methyl- (L) -leucyl residue.

-3'-desoxy-3 'described in the above item (i)
- (C ₁ - ₄ alkoxyimino) MeBmt- residue represented by the formula (IX).

Embedded image (Wherein, R ₉ is C ₁ - ₄ alkyl) alkyl group as R ₉ may be branched or straight chain. Preferably they are linear.

The term N-methyl- (L) -alloisoleucyl (-MealloIle-) used in the above section (iii) is
It means the residue represented by the formula (X).

Embedded image

Preferred cyclosporins of this class are those of the formula (XI)

Embedded image [Wherein, A is -MeBmt-, -dihydro-MeBmt- or the residue described in the above (i), and B is -αAbu-,
-Thr-, -Val- or -Nva-, or additionally A
Is -Ile- when A is -MeBmt- or -dihydro-MeBmt-, and A is -MeBmt- or -dihydro-MeBmt- and B is -αAbu-, -Thr-,-
When Val- or -Nva-, Z is -MeAla-,
When -MeIle-, -MeAlloIle- or -MeLeu-, or A is the residue described in the above item (i), or B is -Ile-, Z is -MeVal- ]
Examples of cyclosporins suitable for use according to the invention include:
It is as follows.

2.1 [3'-desoxy-3'-methoxyimino-MeBmt] ¹ -cyclosporin, 2.2 [Ile] ² -cyclosporin, 2.3 [MeAla] ¹¹ -cyclosporin, 2.4 [MeIle] ¹¹ -cyclosporin, and 2.5 [Me] allo Ile] ¹¹ - cyclosporin, and 2.6 [MeLeu] ¹¹ - cyclosporin.

In this class, the residue at position 2 corresponds to the above (ii)
Preference is given to cyclosporins having the meanings mentioned in the section, especially cyclosporin 2.2. Cyclosporin wherein the residue at position 11 has the meaning as described in the above item (iii), for example, cyclosporin 2.4
And 2.5 are also of particular interest.

Class 3 The above formula (XI) (where Z is -Val- or -MeVal-,
However, when Z is -Val-, A is -MeBmt- or -dihydro-MeBmt-, or Z is -MeVal-.
A is -3'-desoxy-MeBmt-, 3 '
-Desoxy-dihydro-MeBmt-, -N-desmethyl-MeBmt-, -N-desmethyl-dihydro-MeBmt-
-,-3'-desoxy-4'-desmethyl-dihydro-
When MeBmt- or -MeLeu- and Z is -Val-, B is-[alpha] Abu- or -Thr-, and Z is -MeVa.
l- and A are -3'-desoxy-MeBmt-, -3'-
Desoxy-dihydro-MeBmt-, -3'-desoxy-4'-desmethyl-dihydro-MeBmt- or -Me
When Leu-, B is -αAbu-, or Q is -Me
When Val- and A are -N-desmethyl-MeBmt- or -N-desmethyl-dihydro-MeBmt-,
B is -Thr-).

The cyclosporins of this class are as follows: 3.1 [Val] ¹¹ -cyclosporin (also known as cyclosporin E), 3.2 [dihydro-MeBmt] ^1- [Val] ¹¹ -cyclosporin
(Or dihydrocyclosporin E), 3.3 [3'-desoxy-MeBmt] ¹ -cyclosporine (or cyclosporin F), 3.4 [3'-desoxy-dihydro-MeBmt] ¹ -cyclosporine (or dihydrocyclosporin F), 3.5 [N- Desmethyl-MeBmt] ^1- [Thr] ² -cyclosporin (or cyclosporin P), 3.6 [N-desmethyl-dihydro-MeBmt] ^1- [Thr] ²
Cyclosporin (or dihydrocyclosporin P), 3.7 [Thr] ^2- [Val] ¹¹ -cyclosporin (or cyclosporin W), 3.8 [dihydro-MeBmt] ^1- [Thr] ^2- [Val] ¹¹ -cyclosporin (or dihydrocyclosporin) W), 3.9 [3'-desoxy-4'-desmethyl-dihydro-
MeBmt] ¹ -cyclosporin (or cyclosporin Z), 3.10 [MeLeu] ¹ -cyclosporin (or cyclosporin 28).

[0054] (1a) group, particularly (1a ¹⁾ Various acylated cyclosporins of group and (1b) group are known. That is, cyclosporin 1.1, 1.4, 1.8, 1.9, 1.13, 1.14
And 1.34, for example, Traver et al., Helv. Chim. Acta, Vol. 60, 124.
7 pages or less (1977), 65, 1655 pages (19
1982), Vol. 70, p. 13 et seq. (1987), Kobel et al., "European Journal of Applied
Microbiology and Biotechnology "
(Europ. J. Applied Microbiology and Biotechnolog
y), Vol. 14, p. 273 et seq. (1982);
Baltburg and others, "Progress in Allergy" (Pro
gress in Allergy), volume 38, p. 28 or less (1986
) And are described along with their manufacturing methods.

Cyclosporin 1.2, 1.3, 1.5, 1.6, 1.7
And 1.10 are novel and as such form a part of the invention as novel compounds. Cyclosporin 1.2, 1.5-1.
7 and 1.10 represent the corresponding cyclosporins (where the residue at position 1 is -MeBmt- or -dihydro-MeBmt-), for example in analogy to the methods described in the art for the preparation of cyclosporin 1.1. It can be produced by acetylation. Cyclosporine starting materials are known,
Cyclosporins 1.6 and 1.7 are described, for example, in EP-A-0194972. The product, cyclosporin, has the following physical properties.

[Table 1]

In general, the residue at position 2 of the cyclosporin starting material is a β-hydroxy-α-amino acid residue, such as -T
When hr-, the β-OH group at this position is more susceptible to reaction than the 3′-hydroxy of position 1, for example, -MeBmt- or -dihydro-MeBmt-. That is, cyclosporins in which the residue at the 2-position but not the residue at the 1-position is acylated [cyclosporins of the group (1b)]
(Eg, cyclosporin 1.35), or cyclosporins in which both the 1- and 2-position residues are acylated
[In case of cyclosporins 1.13 and 1.14 of group (1a ¹ )]
It can be readily prepared according to or analogous to methods described in the art utilizing the relative differences in reactivity described above. Cyclosporins which are 3'-acylated at position 1 but have a free β-OH group at position 2
In order to produce [for example, in the case of cyclosporin 1.3], it is necessary to first protect the 2-position hydroxy group, then acylate the 1-position residue, and then remove the protecting group. The following example illustrates the general procedure.

Example A Preparation of [3'-O-acetyl-MeBmt] ^1- [Thr] ² -cyclosporin (cyclosporin 1.3) Step 1: Introduction of an O-protecting group at -Thr- ² of [Thr] ² -cyclosporin. 6.09 g of [Thr] ² -cyclosporine was added to 25 ml of anhydrous C
Dissolve in H ₂ Cl ₂ and add 5 ml of ethynyl ether and 0.2
Add ml trifluoroacetic acid. The reaction mixture is stirred for 20 hours at room temperature under a nitrogen atmosphere. 20% cold reaction mixture
Shake with KHCO ₃ , wash with water, extract with CH ₂ Cl ₂ , dry over MgSO ₄ , filter and concentrate. In the resulting foam, 440 g of silica gel (0.04-0.00)
Chromatographic purification using 6), toluene / acetone (2: 1) as eluent, collecting in 500 ml fractions. The product, [Thr-1-methyl-ethoxymethyl ether] ² -cyclosporin, was collected in fraction 2
Recover from 6-29. mp = 76.6 °.

Two steps: [Thr] ² -Cyclosporin -T
hr- ² O-protected acylation. 10.66 g of the product of one step, 106 ml of acetic anhydride and 4
-Combine 1.22 g of dimethylaminopyridine and add 2
Stir for 0 hours and absorb at 40 ° C. in toluene. The residue was dissolved in 400 ml of toluene, once with 250 ml of cold acetic acid, once with 250 ml of H ₂ O and 200 ml of cold 20% K
Extract once with HCO ₃ and once with 250 ml H ₂ O.
The toluene extract was dried over MgSO ₄ , filtered, concentrated,
After vacuum drying, [3'-O-acetyl-MeBmt] ^1- [T
hr-1-methyl-ethoxymethyl ether] ² -cyclosporin is produced.

Three steps: [3'-O-acetyl-MeBmt] ¹
- [Thr] ² - Deprotection of ^-Thr-2 O-protected form of cyclosporin. 11.38 g of the two-stage product, 80 ml of tetrahydrofuran
And 160 ml of 67% acetic acid are stirred under N ₂ at room temperature for 48 hours. The reaction mixture is taken up in toluene, the residue is dissolved in toluene, shaken with 20% KHCO ₃ , washed with H ₂ O, the toluene extract is dried over MgSO ₄ and filtered. The filtrate was concentrated and 440 g of silica gel (0.04-0.
06) and toluene / acetone (3: 1) as eluent
Is used to perform chromatographic purification, collecting in 500 ml fractions. The final product, [3'-O-acetyl-
MeBmt] ^1- [Thr] ² -cyclosporin in fraction 5
Recover from -9. mp = 75.6 °, [α] _D ²⁰ = −27
5.5 ° (c = 0.65 in CHCl ₃ ).

The cyclosporins of the formula (II ') are novel and as such form a part of the invention as novel compounds. Accordingly, the present invention provides in a further aspect a novel group of cyclosporins:
(1a ⁴⁾ those represented by the formula (II ').

The cyclosporins of group (1a ⁴ ) may have been described above for the preparation of cyclosporin 1.3, starting from the corresponding cyclosporin, wherein the residue at position 1 is -MeBmt- or -dihydro-MeBmt-, Alternatively, it can be produced in exactly the same manner as in the production procedure of known cyclosporins 1.1, 1.4 and the like. The required starting materials are known and, together with their preparation, are described, for example, in GB-A-2155936, U.S. Pat. No. 4,639,434 and EP-A-0056678. As described above, cyclosporins represented by the formula (II ′) [where W is a β-hydroxy- (other than β-O-acyl-)-α-amino acid residue] can be obtained, for example, in the same manner as in Example A. , First protecting position 8 and acylating position 1,
It is then prepared by deprotecting position 8.

The cyclosporin 1.15 of the group (1a ⁴ ) has the following characteristic data. [α] _D ²⁰ = −272 ° (c = 1.0, CH
In Cl _3). Cyclosporins in which the 3'-carbon atom of the residue at position 1 is substituted with azidoalkylcarbonyloxy or alkoxycarbonyloxy are also novel and themselves form part of the invention as novel compounds.

The group of cyclosporins of this type includes: (1a ⁵ ) In the formula, the residue at the 1-position is -3′-O- (C
₁ - ₄ azidoalkyl) - carbonyl - or-3'-O-
(C ₁ - ₄ alkoxy) - carbonyl - -MeBmt- or - if a dihydro -MeBmt- residue, i.e., the above formula (I), ACYL ¹ has the formula R ₁ '-CO- or R ₂ -O- CO- (in the formula, R ₁ 'is C ₁ - ₄ alkyl - ₄ azidoalkyl and R ₂ are C ₁₎ is a group represented by. Preferred cyclosporins of this group are those of the above formula (II)
[However, residues of A is the (1a ⁵⁾ above, wherein, B is meaning defined in formula (II), X is -Sar- and Y is -Val-
It is shown by].

The present invention also provides a method for producing cyclosporins of the group (1a ⁵ ), comprising the following steps. a) the corresponding cyclosporin (where the residue at position 1 is -Me
Bmt- or -dihydro-MeBmt-) of formula (XII)
Or (XIII) R ₁ ′ -CO-Q (XII) R ₂ -O-CO-Q (XIII) (wherein R ₁ ′ and R ₂ have the above-mentioned meanings and Q is a leaving group) With the compound to be prepared. Q is preferably halogen or -OH. When Q is a halogen,
The reaction is suitably performed, for example, in the presence of a catalyst (eg, n-butyllithium). When Q is -OH, the reaction is suitably carried out in the presence of an activator. The reaction is carried out, for example, as described in Example B below, preferably from about -100 to -50.
C. at a temperature of .degree.

Embodiment B 1. Production of [3′-O-methoxycarbonyl-MeBmt] ¹ -cyclosporin (cyclosporin 1.11). 3.76 ml of a 1.48 molar solution of n-butyllithium in hexane are added at -78 ° to 0.87 ml of diisopropylamine in tetrahydrofuran.
Stir the whole for a minute. 1.0 g of cyclosporin in 15 ml of dry tetrahydrofuran are added at the same temperature and a further 3
Continue stirring for 0 minutes. 0.326 ml of methyl chloroformate are added and stirring is continued for a further hour at -78 ° C. The temperature is then allowed to warm to room temperature. Add 10 ml of water,
Most of the tetrahydrofuran is removed by concentration. The residue is taken up in 100 ml of ethyl ether and 50 ml of water, the aqueous layer is separated and extracted twice with ethyl ether. The organic extracts are combined, dried over MgSO ₄ and concentrated to dryness. Purify the residue by silica gel chromatography, eluting with ethyl acetate, to give the title compound. [α] _D ²⁰ = -234 ° (c = 1.0, in CHCl ₃ ).

2. [3′-O- (4-azidobutanoyl)-
Production of [MeBmt] ¹ -cyclosporin (cyclosporin 1.12). 7.74 g of γ-azidobutyric acid, 8.6 g of 4-dimethylaminopyridine and 5.11 g of Murayama's reagent (2-chloro-1-methyl-pyridinium iodide) in 12.2 g of 200 ml of anhydrous CH ₂ Cl ₂ . And the reaction mixture is stirred at room temperature for 80 hours. The product obtained was diluted with CH ₂ Cl ₂ and 100 ml of ice-cold 10% NaOH,
Shake with 200 ml of H ₂ O and 200 ml of 10% acetic acid, wash with water and dry the organic phase over Na ₂ SO ₄ . The filtrate is concentrated and dried again. In the product, 1.2 kg of silica gel (0.04-0.06 mm), hexane / acetone (3: 1) as eluent, fraction 20
Up to 250 ml fractions, then hexane /
A chromatographic purification is carried out, eluting with acetone (7: 3) and subsequently collecting in 300 ml fractions. The title compound is recovered from fractions 23-27. [α] _D
²⁰ = -289.8 ° (c = 0.5, in CHCl ₃ ). The cyclosporins of the above group (1a ² ) are also novel and themselves form part of the invention as novel compounds.

The present invention also provides a method for producing cyclosporins of the group (1a ² ), comprising the following steps. b) the corresponding cyclosporin [where the residue at position 1 is of the formula (XI
V)

Embedded image (Wherein, -x-y- and R ₄ have the meanings given for formula (IV)) -8'-C ₁ represented by - ₈ alkoxy - cis -MeBmt- or -8'-C ₁ - ₈ Which is an alkoxy-dihydro-MeBmt-residue], for example, of the formula (X
V)

Embedded image (wherein, R ₅ is C ₁ - ₄ alkyl and Q is a leaving group such as halogen atom and _{a) R 5 -CO-Q (XV} ) by reaction with a compound represented by the above formula (II) wherein A is a residue of formula (XIV), B is as defined for formula (II), X is -Sar- and Y is -Val-. Or

C) The cyclosporin of the group (1a ² )
The residue at the position is the above formula (IV) (where -x-y- is cis-CH
ＣＨCH—, R ₄ has the meaning defined for formula (IV) and A
CYL ¹ is an acyl O-protecting group, eg, acetyl)
3'-O- acyl -8'-C ₁ - ₈ alkoxy - cis -MeB
mt-residue], the residue at position 1 has the formula (XV
I)

Embedded image (Wherein R ₁₀ is an acyl O-protecting group, for example, acetyl) at a -3′-O-acyl-5′-des- (1-propenyl) -5′-formyl-MeBmt-residue. Certain cyclosporins, for example, the above formula (II) (where A is the residue represented by the formula (XVI), B is the meaning defined for the formula (II), X is -Sar- and Y is -Val-) The cyclosporin shown has the formula (XVII)

Embedded image (Wherein, R ₁₁ are each phenyl or C ₁ - is ₄ alkyl, R ₄ have the meanings given for formula (IV)) is reacted with a compound represented by the. The reaction according to the step (c) is
E.g. under anhydrous conditions, e.g. inert solvents or diluents
A standard used to carry out the Wittig reaction in a solvent (e.g., anhydrous benzene, tetrahydrofuran, ethyl ether or toluene) at a temperature of about -80C to 20C, optionally in the presence of a salt (e.g., an alkali metal halide). It can be performed according to the method. The reaction is preferably performed under an inert atmosphere.

The cyclosporin starting materials required for step (c) are the corresponding cyclosporins (where the residue at position 1 is -3′-O-acyl-protected-MeBmt-, for example,
3'-O-acetyl-MeBmt-) d) by ozonolysis and treatment of the immediately obtained ozonolysis product with a mild reducing agent.

The ozonolysis according to reaction step (d) can be carried out according to standard techniques, for example at a temperature of about -90 to about -40 ° C. in the presence of, for example, ethyl acetate or dichloromethane in an ozone generator. A suitable mild reducing agent is dimethyl sulfide. Suitably, the ozonolysis and treatment with a mild reducing agent is performed in a single reaction vessel, for example as described in Example C below.

The starting material for step (d) is a cyclosporin of group (1a ¹ ), which is known or can be prepared as described above. The step (b) is performed, for example, as described above with reference to the literature (1).
a ¹⁾ and a (1b) method for producing a group of cyclosporins may be carried out in a similar manner. Similarly, it has a free β-hydroxy-α-amino acid residue at position 2 such as -Thr- (1a
² ) Cyclosporins of group ² ) can be prepared, for example, by analogous procedures to those described in connection with Example A above, by intermediate protection, followed by deprotection of the β-OH group. The starting material of the above step (b) is

[0072] e) the corresponding cyclosporin (provided that 1-position residues, -8'-C ₁ - ₈ alkoxy - cis -MeBmt-
Or -8'-C ₁ - ₈ alkoxy - dihydro -MeBmt
-Deprotection of the 3'-protected form of the residue) or f) the corresponding cyclosporin, provided that the residue at position 1 is -8 '
-C ₁ - ₈ alkoxy - cis -MeBmt- a free or 3'-O- protected form residues) reduction and, if necessary
It can be produced by performing the step (e).

The appropriate 3′-O in the starting material of step (e)
-Protecting groups include all those known in the peptide chemistry arts, including acyl and ether protecting groups. That is, the starting material of the step (e) includes the product of the step (c). The corresponding starting materials (provided that the 3'-O-protecting group is other than acyl) are prepared analogously to steps (d) and (c) starting from the appropriate 3'-O-protected cyclosporins. Can be done.

The step (e) itself may be carried out according to a procedure commonly used in the field of peptide chemistry, for example, by removing an acetyl protecting group by hydrolysis in the presence of a base (eg, an alkali metal alkoxide or carbonate), or It can be carried out by removing the protected ether group by hydrolytic ether cleavage in the presence of trifluoroacetic acid or HCl. Step (e) is preferably from about -20 to about 20.
C. (for example, Example D below).

Step (f) can also be carried out according to the general method disclosed, for example, in British Patent Specification No. 1567201, by analogous procedures to known methods for the reduction of natural cyclosporins to the corresponding dihydrocyclosporins, for example by catalytic hydrogenation. Can be implemented. The hydrogenation is suitably carried out in the presence of an inert solvent or diluent, for example ethyl acetate or lower aliphatic alkanols (e.g. methanol and isopropanol), a catalyst such as platinum or preferably palladium.
(E.g., palladium on carbon) at atmospheric pressure or slightly above, at a temperature of about 20 to about 30C, at neutral pH.
It is performed under conditions.

The cyclosporin starting material of step (b) is also novel and as such forms a part of the present invention as a novel compound. Accordingly, the present invention provides in another aspect: Residues of Class 4 1-position are represented by the formula (XIV)-8'-alkoxy - cis -MeBmt- or -8'-C ₁ - ₈ alkoxy - Cyclosporin is dihydrofolate -MeBmt- residue. Preferred cyclosporins of this class are those of formula (II) as described in step (b)
It is shown by.

Class 4 cyclosporins also have, in addition to their use as intermediates, for example, the immunosuppressive, anti-inflammatory and antiparasitic activities described below. That is, the cyclosporins have their own uses.

The class 4 cyclosporins, as in the case of the class 1 to 3 cyclosporins, also have the activity of increasing the sensitivity to chemotherapeutic drug therapy or enhancing its effects, in particular chemotherapeutic drug resistance, eg anti- It exhibits activity to counteract resistance to neoplasia or cytostatic chemotherapy. However, class 4 cyclosporins generally have poor suitability for use in the indicated indications, for example, focusing on their inherent immunosuppressive properties.

The cyclosporin starting material of step (c) is also novel and as such forms a part of the present invention as a novel compound. Of course, these starting materials may be deprotected and / or reprotected according to methods known or practiced in the art, for example, for operational, transport or storage purposes. Accordingly, the present invention provides, in yet another aspect, the following.

Class 5 The residue at the 1-position is the free or 3'-O-protected form of -5'-des- (1-propenyl) -5'-formyl-MeBmt-, ie, the formula (XVIII)

Embedded image (Wherein R ₁₂ is hydrogen or an O-protecting group).

A preferred protecting group for R ₁₂ is an acyl O-protecting group, for example, acetyl. Preferred cyclosporins of this class are those of the above formula (II) (where A is of the above formula (XVII)
The residue of I), B is as defined for formula (II) and X is -S
ar- and Y are -Val-).

When R ₁₂ is hydrogen, the residues of formula (XVIII) also exist in cyclic tautomeric forms of formula (XVIII ′).

Embedded image Such tautomers are also within the scope of this invention and are intended to be included, for example, in the above definition of class 5 cyclosporins.

The method for producing the (1a ² ) group and the cyclosporins of classes 4 and 5 will be described below by way of examples. Example C Production of cyclosporins of group (1a ² ). [3'-O-acetyl-8'-methoxy-cis-MeBmt] ¹
Production of cyclosporin (cyclosporin 1.16). 16 g of methoxymethyltriphenylphosphonium bromide in 270 ml of anhydrous tetrahydrofuran in argon
(THF) and cool to -75 ° C. A solution consisting of 5 g of potassium tert-butylate and 200 ml of anhydrous THF is added dropwise over 30 minutes and the mixture is again stirred. After 2 hours, 10 g of [3'-O-acetyl-5'-
Death - (1-propenyl) -5'-formyl -MeBmt] ¹ -
A solution consisting of cyclosporine and 25 ml of THF was added to 2
Add dropwise over 0 minutes. The mixture is stirred at room temperature for 4 hours and then treated with 400 ml of ethyl acetate. The extract is washed with 2N HCl solution, saturated NaHCO ₃ and saturated saline. The filtrate is concentrated and chromatographed on 1.2 kg and 0.44 kg silica gel (0.04-0.063 mm) using CHCl ₃ (10-40%) acetone / ethyl acetate to give the title compound. Can be [α] _D ²⁰ = -2
67 ° (c = 0.53, in CHCl _3).

The cyclosporins 1.17 to 1.21 can be produced in a similar manner. These have the following physical properties:

[Table 2] The starting material for the above process, ie, class 5 cyclosporin, is prepared as follows.

Preparation of [3'-O-acetyl-5'-des- (1-propenyl) -5'-formyl-MeBmt] ¹ -cyclosporin (cyclosporin 5.1). 48.5 g of [3'-O-acetyl- in 70 ml of ethyl acetate
MeBmt] ¹ -Cyclosporin was applied for 45 minutes using a Fischer ozone generator (0.4 atm, flow rate 110 l / h)
Ozone treatment at 70 ° C. The resulting solution is gassed with N ₂ and 9.8 ml of dimethyl sulfide are added. The solution is stirred at room temperature for 2 hours, concentrated by evaporation, washed twice with benzene and dried under high vacuum to give the desired product. It is used directly in subsequent reactions without further purification. [α] _D ²⁰ = −302 ° (c = 1.15, CHCl
₃ ).

The following cyclosporins are produced in the same manner. 5.2 [3'-O-acetyl-5'-des - (1-propenyl) -5'-formyl-MeBmt] ¹ - [O-acetyl -Th
r] ² -cyclosporin, [α] _D ²⁰ = −272.6 ° (c = 0.
504 _{in, CHCl 3), mp = 156-160} °, 5.3 [3'-O- acetyl-5'des - (1-propenyl) -5'-formyl ^{-MeBmt] 1 - [Val] 2} - Ciclosporin, [α] _D ²⁰ = −306 ° (c = 1.03, CHCl
₃ in), mp = 168-170 °, 5.4 [3'-O- acetyl-5'des - (1-propenyl) -5'-formyl ^{-MeBmt] 1 - [Nva] 2} - Ciclosporin, [alpha] _D ²⁰ = −291.76 ° (c = 0.51, CHCl
₃ ).

Example D Preparation of class 4 cyclosporins. Production of [8'-methoxy-cis-MeBmt] ¹ -cyclosporin (cyclosporin 4.1). Cyclosporine 1.16 in 41 ml of methanol / water (5: 1)
4 g (from Example C) are heated with 3.5 g of potassium carbonate and stirred at room temperature. After 21 hours, cool the solution to 10
% Tartaric acid solution by acidification, extracted three times with CHCl _3. The extract is washed with saturated NaHCO ₃ and water, and Na ₂ S
Dry over O ₄ and concentrate. Chromatography on 220 g silica gel with ethyl acetate and ethyl acetate / hexane (100-20%) gives the title compound. [α] _D ²⁰ = −239 ° (c = 0.53, C
In HCl _3).

The following cyclosporins can be obtained in a similar manner. 4.2 [8'-t-butoxy-cis-MeBmt] ¹ -cyclosporin, [α] _D ²⁰ = −228 ° (c = 0.5 in CHCl ₃ ), 4.3 [8′-methoxy-cis-MeBmt] ¹ -[Thr] ² -cyclosporin, [α] _D ²⁰ = −220 ° (c = 0.37, CHCl ₃
Medium), 4.4 [8'-t-butoxy-cis-MeBmt] ^1- [Thr] ^2-
Cyclosporin, [α] _D ²⁰ = −221 ° (c = 0.43, C
In HCl ₃ ), 4.5 [8'-Methoxy-cis-MeBmt] ^1- [Val] ² -cyclosporin, [α] _D ²⁰ = −241 ° (c = 0.54, CHCl ₃₎
Middle), 4.6 [8'-methoxy-cis-MeBmt] ^1- [Nva] ² -cyclosporin, [α] _D ²⁰ = −236 ° (c = 0.634, CH
In Cl _3). Preparation of [8'-methoxy-dihydro-MeBmt] ¹ -cyclosporine (cyclosporin 4.7).

Dissolve 302 mg of cyclosporin 4.1 in 10 ml of absolute ethanol and hydrogenate at room temperature under atmospheric pressure with 5% palladium on carbon. Theoretical amount of hydrogen uptake (5
(0 min), the catalyst is separated by filtration with Hyflo, and the solvent is concentrated under reduced pressure. The residue was treated with hexane-acetone (1: 1).
Chromatography on 70 g silica gel (O = 0.015 mm) using 1) gives the title compound. [α] _D ²⁰ = -229 ° (c = 0.52 in CHCl ₃ ).

The following cyclosporins can be obtained in a similar manner. 4.8 [8'-Methoxy-dihydro-MeBmt] ^1- [Thr] ^2-
Cyclosporine, [α] _D ²⁰ = -217.5 ° (c = 0.3
During _{2, CHCl 3), 4.9 [} 8'- methoxy - dihydro ^{-MeBmt] 1 - [Val] 2} -
Cyclosporin, [α] _D ²⁰ = −223.6 ° (c = 0.5
8 in, CHCl _3), 4.10 [8'- methoxy - dihydro -MeBmt] ¹ - [Nva] ²
-Cyclosporin, [α] _D ²⁰ = -221.5 ° (c = 0.5
94, in CHCl _3).

The cyclosporins of the group (1a ³ ) are also novel and themselves form part of the invention as novel compounds. The present invention also provides a method for producing cyclosporins of the group (1a ³ ), comprising the following steps.

G) corresponding cyclosporin wherein the residue at position 1 is -cis-MeBmt-, -7'-desmethyl-
7'-hydrocarbyl- -MeBmt- or -cis-
A MeBmt-residue (where the hydrocarbyl moiety contains at least 2 carbon atoms), or -7'-desmethyl-
At the 7'-hydrocarbyl-dihydro-MeBmt- residue, wherein the hydrocarbyl moiety has at least 2 carbon atoms and any aliphatic groups or moieties present as or in said hydrocarbyl moiety are saturated. Yes, the residue has the formula (XIX)

Embedded image Wherein -x-y- and R ₆ have the meaning given above for formula (V).
Formula (II) wherein A is a residue of the above formula (XIX), and B is a compound of the formula (I
X is -Sar- and Y is -V
is acylated by, for example, reacting with a compound represented by the formula (XV), or

H) Cyclosporin of group (1a ³ ) [where 1
The residue at the position is -3'-O-acyl-cis-MeBmt- (where the acyl moiety is an acyl O-protecting group), or -7'-desmethyl-7'-hydrocarbyl- -MeBmt.
Or a cis-MeBmt- residue (wherein the hydrocarbyl moiety has at least 2 carbon atoms and the O-acyl moiety is an acyl O-protecting group), said residue having the formula (V) (However, -xy- is cis or trans-
CH = CH-, R ₆ may have the meaning described for formula (V) and ACYL ¹ may be an acyl O-protecting group, for example acetyl). X
VI) cyclosporin, which is a -3'-O-acyl-5'-des- (1-propenyl) -5'-formyl-MeBmt- residue, such as the above formula (II) (where A is the above formula (XVI)
And B is a residue represented by the formula (II);
Is -Sar- and Y is -Val-).

Embedded image (Wherein each R ₁₁ is phenyl or C ₁ - ₄ alkyl and R
₆ 'is a hydrocarbyl having at least 2 carbon atoms) and, if necessary, further isolating the desired reaction product.

Steps (g) and (h) are carried out, for example, in the following Example E
As described later, it can be carried out in exactly the same manner as the steps (b) and (c). The starting material for step (g) is i) the corresponding cyclosporin [where the residue at position 1 is -3 ′
-O-acyl-cis-MeBmt- or -7'-desmethyl-7'-hydrocarbyl- -MeBmt- or -cis-MeBmt- residue, where the hydrocarbyl moiety has at least 2 carbon atoms, This residue is 3 '
-O-protected form] or j) the corresponding cyclosporin wherein the residue at position 1 is -7 '
-Desmethyl-7'-hydrocarbyl- -MeBmt- or -cis-MeBmt- residue, which is in free or 3'-O-protected form] and, if necessary, in step (i) Depending on the implementation, it can be produced in a manner completely similar to the above step (e) or (f).

Suitable 3'-O-protecting groups are the same as described above for steps (e) and (f). When performing step (j), the unsaturated bond of the hydrocarbyl moiety is a group -xy
-React with Alternatively, the cyclosporin starting material (where the residue at position 1 is -cis-MeBmt-) may be used for the overall synthesis of cyclosporins, for example as described in EP-A-0034567 or U.S. Pat. No. 4,369,542. It can be manufactured according to the method.

The cyclosporin starting material of step (g) (where the residue at position 1 is other than -cis-MeBmt-) is also novel and as such forms a part of the present invention as a novel compound. Thus, in another aspect, the invention provides:

Class 6 The residue at the 1-position is a -7'-desmethyl-7'-hydrocarbyl-MeBmt- or -cis-MeBmt- residue (wherein the hydrocarbyl moiety has at least 2 carbon atoms) or -7'-desmethyl-7'-hydrocarbyl-dihydro-MeBmt-residue (provided that the hydrocarbyl moiety has at least 2 carbon atoms and any aliphatic groups or moieties present as or on said hydrocarbyl moiety) Saturated) and the residue is of the formula (XI
X ')

Embedded image (Wherein -xy-is cis or trans -CH = CH
- or -CH ₂ -CH ₂ - and R ₆ "is hydrocarbyl having at least 2 carbon atoms, provided that, -x-y-
Is —CH ₂ —CH ₂ —, wherein R ₆ ″ or any aliphatic groups or moieties present thereon are saturated).

A preferred cyclosporin of this class is
The formula (II) (where A is the residue of the formula (XIX ′), and B is the formula
X is -Sar- and Y is-
Val-). Class 6 cyclosporins also have, for example, the aforementioned immunosuppressive, anti-inflammatory and antiparasitic activities in addition to their use as intermediates. That is, the cyclosporins have their own uses.

Class 6 cyclosporins, as in the case of the above-mentioned class 1-3 cyclosporins, also have an activity of improving the sensitivity to chemotherapeutic drug therapy or enhancing the effect thereof, particularly chemotherapeutic drug resistance, for example, antitumor activity. It exhibits activity to counteract resistance to neoplasia or cytostatic chemotherapy. However, class 6 cyclosporins generally have poor suitability for use in said indications, for example, noting their intrinsic immunosuppressive properties.

The residue at the 1-position is -cis-MeBmt-,
The cyclosporins described and exemplified herein are:
Included within the aforementioned European Patent Publication No. 0034567 and US Pat. No. 4,369,542, but formally new. Hereinafter, the method for producing cyclosporins of group (1a ³ ) and class 6 will be described with reference to examples.

Example E [3'-O-acetyl-7'-desmethyl-7'-phenyl-MeBmt] ¹ -cyclosporine (cyclosporin 1.22) and [3'-O-acetyl-7'-desmethyl-7'- Preparation of phenyl-cis-MeBmt] ¹ -cyclosporin (cyclosporin 1.27). 9.5 g of benzyltriphenylphosphonium chloride are dissolved in 150 ml of dry benzene and reacted with 2.7 g of potassium tert-butylate under reflux for 24 hours on a water separator. After cooling, the suspension is filtered into a new flask under an inert atmosphere. 5 g of cyclosporin 5.1 (see Example C) in 50 ml of benzene are added over 5 minutes. After 21 hours at room temperature, the reaction mixture is poured on ice and the organic phase is extracted with 2N-
Wash with HCl, bicarbonate solution and water, dry over sodium sulfate and concentrate. The residue is chromatographed twice on 440 g of silica gel (0.04-0.063 mm) with chloroform containing 8-30 ppv of acetone. Fraction 2 is cyclosporine 1.27
And fractions 5-15 contain cyclosporin 1.22. Cyclosporins 1.23-1.26 and 1.28-1.34 can be produced in a similar manner. These cyclosporins have the following physical properties.

[0102]

[Table 3]

Example F Preparation of class 6 cyclosporins. Preparation of [7'-desmethyl-7'-phenyl-MeBmt] ¹ -cyclosporin (cyclosporin 6.1). 2.29 g of cyclosporin 1.22 obtained above was
Dissolve in 20 ml of methanol: water (4.5: 1) containing 1.67 g of potassium carbonate. 10% after 20 hours at room temperature
The reaction mixture is poured onto ice containing tartaric acid and extracted with chloroform. The crude extract is chromatographed on 120 g of silica gel (0.04-0.063 mm) with dichloromethane containing acetone (20-30% increase) to give the title compound. [α] _D ²⁰ = -209
° (c = 0.99 in CHCl ₃ ).

The following cyclosporin can be obtained by the same procedure. 6.2 [7'-desmethyl-7'-phenyl-cis-MeBm
t] ¹ -cyclosporin, [α] _D ²⁰ = −236 ° (c = 1, C
During HCl _3), 6.3 [7'- desmethyl-7'-vinyl - cis-MeBmt] ¹
Cyclosporin, [α] _D ²⁰ = −249 ° (c = 1.17,
During CHCl _3), 6.4 [7'- desmethyl-7'-vinyl - cis-MeBmt] ¹
-[Thr] ² -cyclosporin, [α] _D ²⁰ = −248 ° (c =
1.44 in CHCl _3), 6.5 [7'-desmethyl -7 '- (3-methyl -n- butyl)
-Cis-MeBmt] ¹ -cyclosporin, [α] _D ²⁰ = -24
0 ° (c = 1.15 in CHCl ₃ ), 6.6 [7′-desmethyl-7′-n-propyl-cis-Me
Bmt] ¹ -cyclosporin, [α] _D ²⁰ = −220 ° (c = 1.
75, in CHCl ₃₎ 6.7 [7'-desmethyl -7 '- (beta-allyl) - cis -Me
Bmt] ¹ -cyclosporin, [α] _D ²⁰ = −239 ° (c = 0.
97, in CHCl ₃ ), 6.8 [7′-desmethyl-7′-phenyl-MeBmt] ¹ −
[Val] ² -cyclosporin, [α] _D ²⁰ = −208 ° (c =
1.025, in CHCl _3), 6.9 [7'- desmethyl-7'-phenyl - cis -MeBm
t] ^1- [Val] ² -cyclosporin, [α] _D ²⁰ = −240 °
(c = 0.91, in CHCl _3,) 6.10 [7'- desmethyl-7'-vinyl - cis-MeBmt]
^1- [Val] ² -cyclosporin, [α] _D ²⁰ = -254.8 °
(c = 0.48, in CHCl _3), 6.11 [7'- desmethyl-7'-vinyl - cis-MeBmt]
^1- [Nva] ² -cyclosporin, [α] _D ²⁰ = -246.1 °
(c = 0.52, in CHCl _3), 6.12 [7'- desmethyl -7 '- (3-bromo -n- propyl) - cis-MeBmt] ¹ - cyclosporin, [α] _D ²⁰ = -
226 ° (c = 1.44, in CHCl _3).

By deprotection of cyclosporin 1.23, [cis-MeBmt] ¹ -cyclosporin, [α] _D ²⁰ = −235 ° (c
= 0.9 in CHCl ₃ ). [7'-desmethyl-7'-phenyl-dihydro-MeBm
Production of t] ¹ -cyclosporin (cyclosporin 6.13). 796 mg of a mixture of cyclosporin 6.1 and 6.2 are hydrogenated with 100 mg of 10% Pd on carbon in 50 ml of ethanol at room temperature for 6 hours. Filtration on hyflo yields the pure title compound. [α] _D ²⁰ = −210 ° (c = 0.
7, in CHCl _3).

In the same manner, the following cyclosporin is obtained. 6.14 [7'-desmethyl-7'-n-propyl-dihydro-MeBmt] ¹ -cyclosporin, [α] _D ²⁰ = −188 ° (c
= 6.16, in CHCl _3), cyclosporin 6.6 or 6.7
6.15 [7'-desmethyl-7'-ethyl-dihydro-Me
Bmt] ^1- [Thr] ² -cyclosporin, [α] ²⁰ = −224
° (c = 0.4 in CHCl ₃ ), obtained from cyclo D-sporin 6.4, 6.16 [7′-desmethyl-7 ′-(3-methyl-n-butyl) -dihydro-MeBmt] ¹ -cyclosporin, [ α] _D ²⁰
= -222 ° (c = 0.995 in CHCl ₃ ), obtained from cyclosporin 6.5, 6.17 [7′-desmethyl-7′-i-propyl-dihydro-MeBmt] ¹ -cyclosporin, [α] _D ²⁰ = -229 ° (c
= 1.32, in CHCl _3), 6.18 [7'- desmethyl-7'-ethyl - dihydro -Me
Bmt] ^1- [Val] ² -cyclosporin, [α] _D ²⁰ = −23
2.9 ° (c = 0.48 in CHCl ₃ ), cyclosporin 6.1
6.19 [7'-desmethyl-7'-ethyl-dihydro-Me
Bmt] ^1- [Nva] ² -cyclosporin, [α] _D ²⁰ = −22
7.29 ° (c = 0.54 in CHCl ₃ ), cyclosporin
6.20 [7'-desmethyl-7'-phenyl-dihydro-M obtained from 6.11
eBmt] ^1- [Val] ² -cyclosporin, [α] _D ²⁰ = −21
4 ° (c = 1.19 in CHCl ₃ ), obtained from cyclosporin 6.8 or 6.9, 6.21 [7′-desmethyl-7′-ethyl-dihydro-Me
Bmt] ¹ -cyclosporin, [α] _D ²⁰ = −233 ° (c = 1.
15, in CHCl ₃ ), obtained from cyclosporin 6.3.

Obviously, the aforementioned groups (1a ² ) and (1a ³ ) and the cyclosporins of classes 4, 5 and 6 are all novel and structurally related,
For convenience, for example, the residue at position 1 can be included in the unified category including cyclosporins, which is a residue represented by the following formula (XXI), without any problem.

[0108]

Embedded image Wherein (i) Ra is formyl and Rb is as defined for R ₁₂ with respect to formula (XVIII), or (ii) Ra
Is -x'-y'-Rc (wherein -x'-y'-Rc is the formula (IV),
-Xy- as defined for (V), (XIV) and (XIX ')
R _3, are as defined and -x-y-R ₅ or -x-y-R ₅ ")
And Rb is hydrogen or acyl.] Similarly, groups (1a ² ) and (1a ³ ) and classes 4 and 6
Are new and have uses other than as intermediates, for example, when the residue at position 1 is of the above formula (XXI) wherein Ra and Rb are as defined in section (ii) with respect to formula (XXI) Cyclosporins, which are residues represented by the following meaning:

The acyl group as Rb is preferably a group represented by the formula (II)
The group ACYL ¹ as defined for (I) and (IV), in particular (C
₁ - ₄ alkyl) - carbonyl, for example acetyl. Preferred cyclosporins of the category / subcategory are those of the above formula (II) [where A is the residue represented by the above formula (XXI), B is the same as that of the formula (II), X is -Sar- and Y is -Val-].

The various cyclosporins of group (1b) described above, for example, when the residue at position 2 is of the above formula (III ′) (provided that ACY
L ³ is cyclosporin is a residue represented by an acetyl) are known. That is, cyclosporin 1.35 is known and, together with its production method, is described in “Helvetica Simica Acta” (Helv. Chim. Acta), Vol.
247 pages (1977). Other cyclosporins of group (1b) can be produced in a similar manner. However, cyclosporin 1.36 belongs to a completely new type. Thus, yet another group of cyclosporins according to the invention includes the following groups.

(1e) Group i) dicarboxylic acid diester of cyclosporin having β-hydroxy- (L) -α-amino acid residue at position 2, especially ii) cyclosporine having (L) -threonyl residue at position 2 Dicarboxylic acid diesters, and in particular, iii) Formula (XXII)

Embedded image Wherein, R ₁₃ is C ₁ - is ₈ alkylene, CY are each the above formula
(II) (where A is -MeBmt- or -dihydro-Me
Bmt- and B are of the formula (XXIII)

Embedded image (X is -Sar- and Y is -Val-).], A dicarboxylic acid diester represented by the formula:

[0112] Preferably R ₁₃ is C ₁ - a ₄ alkylene. The present invention also provides a process for preparing cyclosporin of the group (1e), comprising the following steps. k) β-O-hemiester of cyclosporin in which the residue at position 2 is a β-hydroxy- (L) -α-amino acid residue, for example, a compound of formula (XXIV)

Embedded image Wherein CY and R ₁₃ are as defined above.
A hemiester or a reactive functional derivative thereof, a cyclosporin in which the residue at the 2-position is a β-hydroxy- (L) -α-amino acid residue, for example, the above formula (II) (where A is-
MeBmt- or -dihydro-MeBmt-, where B is -Thr
-, X is -Sar- and Y is -Val-).

The reaction may suitably be carried out according to the general procedure of Example G below. Example G Preparation of 1,2-ethanedicarboxylic acid [O- (L) threonyl] ² -cyclosporin diester (cyclosporin 1.36). 2-chloro-1-methyl 0.255 g - pyridinium iodide and 0.244g 4-dimethylaminopyridine, in anhydrous CH ₂ in Cl ₂ 1.318g of 5ml at 0 ° C. [(O-
Hemisuccinyl) -Thr] ² -cyclosporin.
1.218 g of [Thr] ² -cyclosporin are added, the reaction mixture is stirred for 3 hours at 0 ° C. and the temperature is raised to 20 ° C. with further stirring. The reaction mixture was diluted with CH ₂ Cl _2, was added to ice and shaken with 0.1 N-NaOH for 10 ml. The mixture was washed twice with H ₂ O, the organic phase was dried over Na ₂ SO _4, filtered, and concentrated. Purification of the crude product by chromatography on silica gel and hexane / acetone (1: 1) as eluent gives the title compound. [α] _D ²⁰ = −209.4 ° (c = 0.5, in CHCl ₃ ).

The [(O-hemisuccinyl) -Thr] ² -cyclosporin required as a starting material is known. For example, International Patent Application No. PCT / EP85 / 00501,
See International Patent Publication No. WO 86/02080, Example 5. Other hemiester starting materials suitable for the preparation of group (1e) cyclosporins may be prepared in a similar manner. Said
Group (1c) cyclosporins are novel and as such form a part of the invention as novel compounds.

The present invention also includes the following steps (1c)
A process for the preparation of a group of cyclosporins is provided. l) oxidize cyclosporin in which the residue at position 1 is 3'-hydroxy substituted, for example -MeBmt-. The step (1) can be carried out, for example, by reacting with, for example, N-chloro-succinimide / dimethyl sulfide at a temperature of, for example, -30 ° to 10 ° C. according to the procedure of Example H below.

Example H Preparation of [3'-desoxy-3'-oxo-MeBmt] ¹ -cyclosporin (cyclosporin 1.37). 8.48 ml of dimethyl sulfide are added at 0 ° C. with 12.8 g of N
To a solution consisting of chlorosuccinimide and 400 ml of toluene. The mixture was stirred at 0 ° C. for 5 hours,
Cool to 12 ° C. and add 9.62 g of “cyclosporin” in 40 ml of toluene. The resulting suspension was cooled at -10 ° C for 1.
Stir for 5 hours and 1.0 hours at -10 to -5 ° C.
When 19.4 g of triethylamine are added, a pale precipitate forms there. The mixture was stirred at 0 ° C. for 5 hours,
Dilute with ml of ethyl ether. 192 ml of 1N-HC
Add l, cool and extract. The organic phase was washed with 500 ml H ₂ O
And twice with 250 ml of cold 10% NaHCO ₃ , washing twice with 500 ml of H ₂ O, 250 ml of 10% tartaric acid and again with H ₂ O. The acidic and basic aqueous solutions are re-extracted with 2.times.500 ml of ethyl ether. The organic phases are combined, dried over Na ₂ SO ₄ and concentrated to dryness at 40 ° C.
Purify the residue by chromatography using 300 g silica gel and saturated aqueous ethyl acetate to give the title compound. [α] _D ²⁰ = -241.3 ° (c = 0.
531 in CHCl ₃ ) and -169.5 ° (c = 0.5,
CH in ₃ OH).

Cyclosporins 1.38 and 1.39 can be prepared in a similar manner.

[Table 4] The cyclosporins of the group (1d) are also novel and themselves form part of the invention as novel compounds.

The present invention also includes the following steps (1d)
A process for the preparation of a group of cyclosporins is provided. m) oxidize, for example selectively oxidize, cyclosporine in which the residue at position 2 is a β-hydroxy-α-amino acid residue, for example a (L) -threonyl residue. Step (m) can be performed in exactly the same manner as in step (l) above, for example, according to the general procedure of Example I below.

Example I Preparation of [α-methylketo-Gly] ² -cyclosporin (cyclosporin 1.40). 484 mg of dimethyl sulfide are added at 0 ° C. to 868 mg of N-chlorosuccinimide in 26 ml of toluene. 1
After stirring at 0 ° C for 0 minutes, the suspension was cooled to -30 ° C,
1584 mg of [Thr] ² -cyclosporin in 6.5 ml are added. The reaction mixture was stirred at 26-30 ° C for 1 hour, and 1.
81 ml of triethylamine are added. The cooling is released and after a further 5 minutes 25 ml of ethyl ether are added. The reaction mixture was poured into 150 ml ice water / 10 ml 1N HCl,
Stir and add another 100 ml of ethyl ether.
The organic phase is washed three times with 150 ml of ice-water and extracted twice with ethyl ether. The combined organic phases are dried over Na ₂ SO ₄ and concentrated. The residue was dissolved in 10 ml of CH ₂ Cl ₂ ,
Filter through Hyflo, dilute with hexane and concentrate to give the title compound. [α] ²⁰ = −229 ° (c =
1.0, in CHClD ₃ ) and -184.8 ° (c = 1.0,
CH in ₃ OH). Cyclosporin 1.41 can be produced in a similar manner. [α] _D ²⁰ = −226.7 ° (c = 1.0, CHCl ₃
During).

The class 2 (i) and class 2 (ii) cyclosporins described above are also novel and themselves form part of the invention as novel compounds. Further, the present invention also provides a process for producing class 2 (i) and 2 (ii) cyclosporins, comprising the following steps.

M) When preparing a cyclosporin of class 2 (i), a cyclosporin in which the 3'-carbon atom of the residue at position 1 is oxo substituted, for example, the residue at position 1 is -3'-desoxy-3 '- cyclosporin is oxo MeBmt- C ₁ - or ₄ is reacted with an alkoxyamine, or n) when preparing the cyclosporin class 2 (ii), the 2-position (L) - cyclosporin having isoleucyl residues Open-chain peptide containing the sequence (however, this open-chain peptide is
Starting at the residue corresponding to residue 8 of the cyclosporin as the N-terminus and ending with the residue corresponding to residue 7 at the cyclosporine as the C-terminus), for example, closing an open-chain peptide comprising the following sequence: I do.

Embedded image

[0122] The above (m) step, for example in accordance with the general procedure of the following examples J, in the presence of a base (e.g., pyridine) at a temperature of, for example, about -10 ° ~ about 80 ° C., C ₁ which is selected - _The reaction can be carried out by reacting the ₄ alkoxyamine with a suitably acid addition salt form. The step (n) can be carried out, for example, in U.S. Pat.
It can be carried out in exactly the same way as described in US Pat. No. 456 for general synthesis of cyclosporins. That is, cyclosporin 2.2 is, for example, BOC-αAbu-O in step (a) of Example 1 of the patent / patent publication.
It is prepared by replacing H with BOC-Ile-OH and then proceeding in a straightforward manner steps (b) to (x) described in the above mentioned patent / patent publication. Cyclosporin 2.2 itself has the following characteristic data. Melting point 151 ° C., [α] _D ²⁰ = −224 ° (c = 1.0, CHC
l ₃ ).

Example J [3'-desoxy-3'-methoxyimino-MeBmt] ^1-
Production of cyclosporin (cyclosporin 2.1). 600 mg of the product of Example H and 835 mg of methoxylamine hydrochloride in 2.5 ml of ethanol / 2.5 ml of pyridine
Is stirred at 50 ° C. for 14 hours under an argon atmosphere. The pyridine is concentrated off and the residue is filtered. The filtrate is CH ₂ Cl
Absorb ₂ and wash with saline. The brine is extracted several times with CH ₂ Cl ₂ , the combined organic phases are washed again with brine, dried and concentrated. Purify the residue by chromatography to give the title compound. [α] _D ²⁰ = -214
° (c = 1.09 in CHCl ₃ ).

It is clear that the above-mentioned cyclosporins of group (1c) and class 2 (i) are all novel and have uses other than as intermediates, for example the residue at position 1 Is the formula (XXV)

Embedded image Wherein R ₄ is oxygen or R ₁₅ N = (where R ₁₅ is C _{1-
4 is} alkoxyl), which is a residue of cyclosporine.

Class 2 (iii) cyclosporins are known. That is, cyclosporins 2.3 to 2.6 are described in, for example, Wenger et al., "Transplantation Proc."
(December), pages 213 et seq. (1986). These cyclosporins and other cyclosporins of class 2 (iii) are described, for example, in US Pat.
No. 31 and EP-A-0098456, according to the general procedure for the overall synthesis of cyclosporins, for example in step (g) of Example 1 with H-MeVal-
Bzl is converted to H-MeIle-Bzl or H-MeAllo-Ile-Bzl.
And can be produced by substituting Cyclosporin 2.
3 to 2.5 have the following characteristic data.

[Table 5]

Class 3 cyclosporin 3.1-3.5, 3.7
And 3.9 are known [eg, Traver et al., "Helv. Chim. Acta", 65.
Volume, p. 1655 et seq. (1982) and 70, p. 13 et seq. (1987)].

Cyclosporins 3.6, 3.8 and 3.10 are novel and as such form a part of the present invention as novel compounds. Cyclosporins 3.6 and 3.8 can be produced in exactly the same manner as used for the production of other known dihydrocyclosporins, for example as described above for step (f).

Cyclosporin 3.10 is available as a small intermediate metabolite fraction from the fermentation broth used in the production of [(D) Ser] ⁸ -cyclosporin, for example as described in Example K below (EP-A-0056678). See, for example, "Helvetica Simica Actor" (Helv. Ch.
im. Acta) 65, 1655 et seq. (1982), and can be prepared according to the general procedure for the separation of small intermediate metabolites of cyclosporin.

Example K Production of [MeLeu] ¹ -cyclosporin (cyclosporin 3.10). The biomass obtained after the cultivation according to Example 3 of EP-A-0056678 is spun down in a purification separator (Westphalia) and the liquid phase (culture filtrate) is extracted three times with an equal volume of ethyl acetate each time. The ethyl acetate extract is concentrated under vacuum. The solid phase (mycelium) is treated with methanol and homogenized, and the solid and liquid phases are separated by a clarifying separator. This extraction process is repeated twice using 90% methanol. The methanolic extracts are combined, water is added and the extracts are concentrated in vacuo. The remaining aqueous extract concentrate is extracted three times with an equal volume of ethyl acetate and the ethyl acetate extract is concentrated under vacuum. Both crude extracts (from culture filtrate and mycelium)
The silica gel (0.040-0.063 mm) and H ₂ O
-Chromatography using saturated ethyl acetate. The first eluting fraction contains mainly [Val] ² -cyclosporin (cyclosporin D), the later fraction contains mainly [MeLeu] ¹ -cyclosporin (cyclosporin 3.10) and “cyclosporin” (cyclosporin A). The peak fraction is again chromatographed on silica gel (0.020-0.045 mm) and acetone / hexane (1: 1) as eluent. Lichoprep® RP-18 (0.04
0-0.063Mm) and methanol as eluent / H ₂
O (85:15) followed by silica gel (0.020-0.04).
Rechromatography of the fraction containing [MeLeu] ¹ -cyclosporin using 5 mm) and H ₂ O-saturated ethyl acetate as eluent gives the title compound. mp = 142-148 ° C., [α] _D ²⁰ = −303 ° (c =
0.54, in CHCl _3). The physical properties of cyclosporin 3.6 are as follows. mp = 180-182 ° C., [α] _D ²⁰ =
-211 ° (c = 0.64, in CHCl _3).

[0130] As already indicated, this invention, class ^{1 [(1a 1) ~ (} 1a 5) containing group and (1b) ~ (1e) group cyclosporins, cyclosporins of 2 and 3, in particular already Individually named these classes of individual cyclosporins may be used in other chemotherapeutic medications, especially in antimicrobial (e.g., antibacterial, antiviral, antifungal or antigenic animals)
It has been found that chemotherapy and especially anti-cancer or anti-tumor (eg, antineoplastic or cytostatic) chemotherapy can be enhanced or enhanced in potency, or can be enhanced or increased in sensitivity thereto. Thus, they are useful as a means of reducing overall drug toxicity, more particularly as a means of counteracting or reducing resistance, including both innate and acquired resistance to chemotherapy, e.g., antineoplastic or cell proliferation In the case of inhibitory drug therapy, it is useful, for example, as a means of lowering conventional chemotherapy dose levels.

Example 1 Use in anti-neoplastic / inhibition of cell proliferation, restoration of sensitivity to antitumor drug substances (in vitro-1). Twentyman et al., "The British Journal of Cancer" (Br. J. Cancer), 54, 253 (198).
6) according to the method described in Daunorubicin (D
R), vincristine (VC), adriamycin (A
M), etoposide (ET), tenoposide (TE) and colchicine (CC)
(CTDS) resistant to one or more of
For example, a cancer cell line (CCL) derived from human small cell carcinoma of the lung
Grow.

By measuring the inhibition of cell growth between successive CTDS exposures, the resistant subline (CCL-
R) sensitivity compared to that of the parental sensitivity line (CCL-S), eg, in the case of the DR-resistant line (CCL-DRR), CCL-DRS and CCL in the presence of DR originally contained in the growth medium -Compare DRR line growth. To do so, cell proliferation is measured by counting cells using an electronic cell counter, and counting should be performed at a point near the end of the exponential growth phase. For CCL-R line, IC ₈₀ [Non-CTD
The drug concentration required to reduce the final cell number to 20% of the S (eg, DR) treated control, eg, the DR concentration] is> 80 times, preferably >>, as compared to the parental CCL-S line. Select one that is 100 times larger.

Next, the CTD of the selected CCL-R line in the presence and absence of the test cyclosporin
Sensitivity to S (eg, DR) is measured using cell counting as a measure of proliferation as described above. For this, the cells are initially cultured in the presence of various concentrations of both CTDS and the test cyclosporin substance. In the screening, a concentration is selected for the latter that does not itself cause a significant decrease in proliferation. CC in the presence of various concentrations of cyclosporine in the absence of CTDS
Appropriate concentrations are established by culturing LS and CCL-R. Cyclosporins are usually 0.01 to 5
0, especially 0.1 to 10 μg / ml, for example 0.01, 0.0
2, 0.05, 0.1, 0.2, 0.5, 1.0, 2.0,
Tested at concentrations of 5.0, 10.0 and 50 μg / ml. The concentration of CTDS (eg, DR) required for 50% inhibition of cell proliferation in the absence of test cyclosporin (IC ₅₀ −
The percentage obtained by comparing (CS) with the concentration obtained in the presence of the test cyclosporin (IC ₅₀ + CS) is defined as a measure of the enhanced sensitivity of the CCL-R line to CTDS induced by cyclosporine. The stability of the CCL-R line used is ensured by cross-checking its susceptibility to CTDS with already established issues.

In the above test model, cyclosporins of classes 1 to 3, in particular the cyclosporins specifically listed, are capable of reacting with CTDS (eg DR, VC, AM, etc.) at the above concentrations, especially at a concentration of about 10 μg / ml or less. Effective for increasing sensitivity. High concentration, eg about 50μg
/ Ml, in certain cases CCL-S and CCL
Inhibition of growth of both -R lines is induced by test cyclosporin in the absence of CTDS, but if this occurs,
This phenomenon is generally less pronounced on the CCL-R line than on the CCL-S line. More importantly, the test cyclosporin was tested in the CCL-R line at concentrations that did not affect cell proliferation in the absence of CTDS.
It was found to be effective in increasing the sensitivity to DS.

[0135] That is, in a series of tests using the AM resistant human small lung carcinoma cell line as well as cyclosporin 2.4,1.1 and 1.32, established AM sensitization ratios (AMIC ₅₀ -
CS / AM IC ₅₀ + CS) is as follows (*).

[Table 6] [* Twentyman, MRC Clinical Oncology and Radiotherapeutics.
Units (Hills Road, Cambridge, UK)
Research reports and data obtained from the National Institute of Technology, British Journal of Cancer (Br. J. Cance)
r) 57, 254-258 (1988)]

Example 2 Use in antineoplastic / inhibition of cell growth, restoration of sensitivity to antitumor drug substances (in vitro-2). This test is described in, for example, Ring et al., “Journal of Cellular Physiology” (J. Cell. Physiol.), 83, 10
3-116 (1974) and Bege-Hansen et al.
"Journal of Cellular Physiology" (J.
Cell. Physiol.) 88, 23-32 (1976), and can be performed using any suitable drug resistant and control (parent) cell lines. The particular clones selected are those that have multiple drug resistance (e.g.,
Colchicine resistant) line CHR (subclone C5S3.
2) and the parental sensitivity line AUX B1 (subclone AB1 S11) [Reference, Ring et al., Supra and Giuliano et al., Biochim. Biophysica Acta, 455, 152]. -1
62 (1976), Carlsen et al., Biochim. Biophysica Acta (Biochim. Biophys. Acta),
455, 900-912 (1976), Lalande et al., "Proceedings of the National Academy of Sciences" (Proc. Natl. Acad.
Sci.), 78 (1), 363-367 (1981),
"Science", 221 volumes, 1
285-1288 (1983), Cartner et al., Nature, 316, 820-823 (1
985), Riordan et al., Nature, 3
16, 817-819 (1985), van der Brick et al., "Mol. Cell. Biol" (Mol. Cell. Bi.).
ol.), 6, 1671-1678 (1968), Endicort et al., "Mol. Cell. Biol" (Mol. Cell. Bi.
ol.), 7, 4075-4081 (1987), Duchar et al., "Mol. Cell. Biol."
l.), 7, 718-724 (1987) and Gerlach et al., Nature, 324, 485.
-489 (1986)].

(L) -asparagine 0.02 mg / ml, ME
M vitamin (IX), penicillin-streptomycin 10
0 UT / ml, 2 mmol of (L) -glutamine and 10%
Cell lines are grown in αMEM medium supplemented with heat inactivated fetal serum. The assay is performed using a 96-well plate. 50 μl of colchicine solution was added to the culture medium to prepare the same three sets, and the resistance line (RL) was 3
0-10-3-1-0.3-0.1-0 μg / ml and for the sensitivity line (SL) 0.3-0.1-0.03-
A final concentration of 0.01-0.003-0.001-0 μg / ml is achieved. If necessary, ie if the test cyclosporine reduces the IC ₅₀ response to colchicine very significantly, a further extension of the dose range is carried out.

Test cyclosporine was added to anhydrous C ₂ H ₅ OH at 1
Dissolve at a rate of mg / ml. 0.1% of each test cyclosporin
And 1.0 μg / ml at a concentration of 1.0 μg / ml, and the control is treated with the corresponding C ₂ H ₅ OH solvent dilution. Test cyclosporine or control is added to each well (50 μl) and mixed with the existing colchicine solution. S
For L, 4 × 10 ³ cells / ml (400 cells / well)
And for RL add 100 μl of cell suspension at 8 × 10 ³ cells / ml.

3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyltetrazolium bromide (MT
T) using a colorimetric assay [Mossman, Journal
Of Immunological Methods ”(J. Immunol. Meth
ods), 65, pp. 55-63 (1983)]. First, 100 μl of cell-free supernatant is removed, then 10 μl of a 5 mg / ml MTT solution in RPMI 1640 medium (Gibco) is added to each plate and the plates are incubated at 37 ° C. for 3 hours. Then 10
0 μl of butanol / isopropanol / 1N HCl (1
92:96:12 ml) is added to each plate and the plates are shaken until completely dissolved. Optical density (OD) of 540
Read in nm.

The cell growth range (measured by MTT-dependent OD) was expressed as a function of colchicine concentration, and a dose response curve was drawn for each test cyclosporin, with colchicine IC
_{The 50} (CIC ₅₀ ) (ie, the concentration of colchicine required for 50% growth inhibition) is determined. 0.1 and 1.0 μg /
The enhancement of colchicine sensitivity induced by each test cyclosporine at both ml concentrations is calculated as follows.

(Equation 1) Test cyclosporine can affect colchicine sensitivity in both RL and SL.

The relative effects on the two lines can be calculated as follows.

(Equation 2) Class 1-3 cyclosporins enhance the sensitivity of RL to colchicine at the above concentrations. The enhancement of the sensitivity of SL is generally inferior to that of RL, but is usually observed at equal concentrations. In applications according to the present invention, the above-mentioned relatively resistant cyclosporins are generally considered more suitable. In a series of experiments, the following enhancements in sensitivity to colchicine were determined, for example, by RL
And SL.

[Table 7]

Example 3 Application in vivo (anti-neoplastic / cytostatic, restoring sensitivity to antitumor drug substances). Drug substance DR, V
The Ehrlich ascites cancer (EA) subline, which is resistant to C, AM, ET, TE or CC, is described in Slater et al., Journal of Clinical Investigation, J. Clin. Invest. Page 1131 (1982
Year), the next generation of EA cells (BA
It is grown by serial passage into (LB) / c host mice. For this reason, the drug substance was administered at a dose of 0.2-0.5 mg / kg over 5 doses starting 24 hours after inoculation of the host mice with 0.2 ml of undiluted malignant ascites taken from the animal before death. Administer intraperitoneally daily.

To make the test accurate, the host mice were given the EA of the resistant EA-R or susceptible (parent) EA-S as described above.
Administer the line. The mice to which EA-R has been administered are divided into the following groups. 1. 1. No drug / no cyclosporine, 2. Drug substance treatment / no cyclosporine, 3. No drug / test cyclosporine, Drug substance + test cyclosporin.

The antineoplastic drug substance is administered at the dose adopted in the production EA-R line. Starting 24 hours after inoculation with EA-R, cyclosporine test substance is injected intraperitoneally (in ethanol / olive oil) in 5 divided daily doses from 1 to 80 mg / kg, in particular 5 or 25 or 40 mg.
/ Kg of total test dose. The average survival time in groups 2, 3 and 4 is compared to group 1 as a measure of the effect of the applied treatment.

The results obtained show that there is no significant difference in the mean survival time between the first, second and third groups. In the case of group 4, administration of class 1-3 cyclosporine at the above doses results in a substantial increase in survival time compared to both groups 1 and 2 (eg, on the order of 2-3 times or more). ) Is observed. Use class 1-3 cyclosporins in a test model of comparable design (eg, in vitro) or use drug-resistant viral strains, antibiotic (eg, penicillin) -resistant bacterial strains, antifungal-resistant fungal strains and drugs Resistant protozoan strains, such as malaria pathogen strains, such as Plasmodium falciparum exhibiting acquired chemotherapeutic antimalarial drug resistance
Equivalent results can be obtained using test animals infected with a natural substrain of Arum). The use of the class 1 to 3 cyclosporins according to the invention can also be demonstrated in clinical trials, for example in preliminary trials carried out as follows.

Clinical Test 1 Patients diagnosed as exhibiting an increase in malignant cancer and subject to anti-neoplastic / cytostatic drug therapy should be treated
(Male and female). For each subject who initiated the study, a detailed record listing the medical history, medical condition, rate of establishment of disease progression, and estimated prognosis is presented. For the type and extent of cancer, the attending physician will determine the type of anti-neoplastic / cytostatic drug therapy to apply and the estimated dose rate required without cyclosporine therapy.

The test cyclosporine was administered in an amount of 1.0 to 20.0 mg.
/ Kg / day orally or 0.5-5.0 mg /
Administer parenterally (eg, intravenously or intramuscularly) at a dose rate of kg / day. If the condition permits, treatment with the test cyclosporin is initiated at least one or two days, preferably 10-14 days prior to commencing anti-neoplastic therapy, in order to augment the ongoing therapeutic dose level. In such cases,
Starting cyclosporine dose is the lower of the above dose range
(Eg, on the order of 1.0-5.0 mg / kg / day for the oral route, or 0.5-1.5 mg / kg / day for the parenteral route)
Higher dose levels (eg, on the order of days) up to 5.0-15.0 or 20.0 mg / kg / day for the oral route, or 2.0-5.0 mg / kg for the parenteral route. kg
/ Day order), and the attending physician will determine the correct intake, taking into account the subject's condition at the start of the study. In some cases, it may be necessary to start and stop cyclosporine and anti-neoplastic / cytostatic therapy simultaneously, but it is preferable to increase daily from a low starting cyclosporine dose to the indicated maximum dose.

Initial anti-neoplastic / cytostatic therapy is initiated at approximately one third of the estimated dose rate required without cyclosporine therapy, but at the discretion of the attending physician, A selection is made. The anti-neoplastic / cytostatic dose is increased as required by the observed response. During the course of the study, cyclosporine and antineoplastic /
Monitor all relevant clinical parameters, including the rate of administration of the cytostatic agent. Subjects are monitored for control / reduced tumor growth / occurrence of metastases and possible tumor regression. Reports of medical condition and possible prognosis will be submitted at intervals throughout the study.

When the test results were evaluated, the participating subjects were:
Restriction of tumor growth or reduction of tumor regression and metastasis at dose rates less than the anti-neoplastic / cytostatic drug dose rate estimated to be necessary to achieve an equivalent effect without cyclosporine therapy It has been shown to exhibit effective control or improvement of the associated symptoms. Reduced incidence of anti-neoplastic / cytostatic drug resistance and toxic side effects to administered anti-neoplastic / cytostatic drug therapy, as compared to reports for the group receiving only anti-neoplastic / cytostatic drug therapy Has been reported.

Clinical Trial 2 Subjects are selected from hospitalized or outpatient (male and female) patients presenting with an increase in all types of terminal malignancy. Subjects selected for the study already received maximal anti-neoplastic / cytostatic drug therapy and were generally located at the end of multi-drug treatment,
Patients who exhibit tumor growth recurrence / metastasis, ie, whose cancer is clear evidence of multidrug resistance. A detailed report listing each of the subjects participating in the study, in particular the medications, medical histories and recent medical conditions applied during the course of 6-18 months ago to the present, e.g. the estimated speed of disease progression and the estimated prognosis. Submit.

For the purposes of this study, the anticipated dose and schedule of antineoplastic / cytostatic drug therapy will be continued for participating patients, but the selected therapy will be applied at the time of drug resistance confirmation and study initiation. Things. Anti-neoplastic drug therapy is continued throughout the study at pre-resistance-measured levels and schedule. An oral dosage form of about 1 to about 20, eg, about 5 to about 15 mg / kg / day, or a daily dose rate of about 0.5 to about 7.5, eg, about 2.0 to about 5.0 mg / kg.
Chemotherapy is supplemented by administration of the test cyclosporine by the parenteral (eg, intravenous) route in kg / day. In some patients, cyclosporine therapy may be initiated 1-14 days prior to the onset of anti-neoplastic therapy, and will be continued for the entire treatment period. On the other hand, it may be necessary to start and end cyclosporine and anti-neoplastic therapy simultaneously.

Subjects are monitored for the development of tumor growth shrinkage / metastasis and possible regression of the tumor. Reports of medical conditions and possible prognosis at the time of the study will be submitted at intervals throughout the study. If no improvement or worsening of the condition is reported within a reasonable period of time, the basic antineoplastic / cytostatic drug therapy will be modified at the discretion of the attending physician for another previously applied therapy. During the study, all relevant clinical parameters are monitored throughout, including serum levels and clearance rates, particularly of anti-neoplastic / cytostatic drugs and cyclosporine.

In evaluating the study reports, participants showed a marked improvement in the condition associated with suppression of tumor growth or reduction of tumor regression and metastasis following induction of cyclosporine therapy, followed by an improvement in disease prognosis. It turns out that it shows. Thus, class 1-3 cyclosporins are useful as treatments for disease states that exhibit acquired resistance to chemotherapeutic drug treatment or as adjuvants for chemotherapeutic drug treatment.

Thus, the present invention provides the following. 1.1 a method of improving or enhancing the effect of, or increasing the sensitivity to, chemotherapeutic drug therapy, or 1.2 a method of reducing the proportion of an effective chemotherapeutic agent dose, wherein said class 1-3 cyclosporine is in a subject in need of treatment. Or 2.1 a method of treating a disease state that exhibits or is characterized by acquired, induced or congenital resistance to chemotherapeutic treatment; or 2.2 acquired, induced or congenital to said treatment To promote or ameliorate chemotherapeutic treatment of disease states that exhibit or are characterized by, or 2.3 to counteract or reduce acquired, induced, or innate resistance to chemotherapeutic treatment; or 2.4 chemotherapeutic treatment A method of restoring susceptibility to
A method comprising administering to a subject in need thereof an effective amount of a cyclosporin belonging to any one of Classes 1-3 above.

On the other hand, the present invention provides the following. 3. A method of chemotherapeutic treatment comprising administering to a subject in need thereof a cyclosporine belonging to any one of Classes 1 to 3 as an adjuvant treatment for said drug substance together with a suitable chemotherapeutic drug substance. The present invention also provides the following. 4. 4. a cyclosporin belonging to any one of the above classes 1 to 3, which is used in the method according to any one of the above 1.1, 1.2, 2.1, 2.2, 2.3, 2.4 or 3; 4. A cyclosporin belonging to any one of Classes 1 to 3, which is used for producing a pharmaceutical composition used in the method according to any one of 1.1, 1.2, 2.1, 2.2, 2.3, 2.4 or 3.

None of the class 1-3 cyclosporins has been previously proposed or recommended for pharmaceutical or therapeutic use, and certain class 1-3 cyclosporins are in fact new compounds or completely new types of cyclosporins. Accordingly, the present invention also provides the following. 6. Individual cyclosporins which are used as medicaments, for example, which have already been shown to be novel, as well as (1
^{a 2), (1a 3)} , (1a 4), (1a 5), (1d), specifically above classes containing cyclosporins (1e) group and class 2 (i) and 2 (iii) 1-3 cyclosporins.

The particular condition / form of the chemotherapeutic treatment to which the method according to the invention is applied is one or more antimicrobial or antibiotic agents, such as antiviral, antibacterial, antifungal or antigenic animal drug substances Microorganisms, including strains that are resistant to, such as viruses, bacteria, fungi or protozoa, include conditions caused by infection. The method of this invention may be used to reduce or counteract, for example, the growth of tumors, the occurrence of metastases, etc., as one or more anti-cancer chemotherapeutic agents, such as antineoplastic or cytostatic agents, such as anthracyclines Or exhibiting induced or acquired resistance to a vinca alkaloid drug substance or a specific drug substance, daunorubicin, vincristine, adriamycin, etoposide, tenoposide and / or colchicine, in particular cancer, such as a carcinoma, sarcoma or other tumor or malignant tumor Applicable to such treatment.

Preferred cyclosporins for use according to the invention are those having relatively low immunosuppressive activity, for example, those which exhibit substantially no immunosuppressive activity at defined dose levels,
Or, it exhibits immunosuppressive activity substantially lower than that of "cyclosporin", for example, of the order of <50%. Particular cyclosporins suitable for use according to the method of the present invention are the cyclosporins specifically mentioned above as Classes 1-3.

Of course, the dose of cyclosporin to be used when practicing the above method will depend, for example, on the condition to be treated.
(Eg, the type of disease and the nature of the resistance), the particular cyclosporin used, the effect desired and the mode of administration. However, in general, once a day or 2-4
Oral administration in doses in the order of 1 to 20 to 50 mg / kg / day in divided doses, for example in the order of 5 to 10 mg / kg / day, or 0.5 to 7.5 to 10
parenteral in doses on the order of mg / kg / day, for example on the order of 1.5 or 2.0 to 5.0 mg / kg / day (eg,
Intravenous, eg, intravenous, infusion or infusion) administration will give satisfactory results.

That is, a daily dose suitable for a patient is 50
On the order of 1000 to 2500 mg (oral), for example on the order of 250 to 500/600 mg (oral), or 2
5.0 to 375.0 to 500 mg order (intravenous),
For example, the order of 75.0-100 / 250mg (intravenous)
Is within. Alternatively and more preferably, dosages can be distributed in a patient-specific manner that provides a predetermined trough blood concentration, for example, as measured by the RIA (radioimmunoassay) technique. That is, by adjusting the patient's dosage, a constant continuous trough blood concentration as measured by RIA on the order of 50 or 150 to 500 or 1000 ng / ml can be achieved. That is, in this case, the administration method similar to the administration method used recently in the usual "cyclosporin" immunosuppressive therapy is performed.

In practicing the method according to the present invention, a pharmaceutical composition suitable for oral administration of the above-mentioned class 1-3 cyclosporins can be produced, for example, in the following manner.

TABLE 8 Ingredient Relative amount (g) i) Labrafil (Labrafil) M 1944 CS 0.99 ii) cyclosporin (e.g., consequent Rosuporin 1.1, 1.2, 1.3, 25 1.11, or 1.36) (dry weight value) iii) ethanol (anhydrous) 50 iv) Olive oil about 237 (appropriate amount) Total about 462.0

All components are weighed directly in a stirred vessel. Components (i) and (iii) are weighed first. Subsequently, component (ii) is added with continuous stirring until dissolved. Then the ingredients
(iv) is added with stirring for a further 10 minutes. The solution obtained is brought to about 0.5 to 0.8 bar at a Germanic preflow.
Filter through a filter (400 μm),
(Rexo) Fill the bottle. The jar is sealed with a rubber cork and cap. While supplying nitrogen in all stages (0.25
Bar) All steps are carried out at room temperature under water-free conditions. A bottled solution suitable for oral administration is 50 mg of cyclosporine
(Dry weight) / ml. Alternatively, the solution can be filled with gelatin, for example a hard or soft gelatin capsule, as a means of masking taste.

The class 4 and class 6 cyclosporins, in particular the classically named cyclosporins, also exhibit pharmacological activity. In particular, they exhibit immunosuppressive, anti-inflammatory and antiparasitic activity, for example as shown in the in vitro and in vivo tests described below. The cyclosporins also have activity on experimental allergic encephalomyelitis (EAE) in rats as described below.

Example 4 Immunosuppressive activity 4.1 Local in vitro hemolysis in gel [Michel and Dutton, "Journal of Experimental Medicine" (J. Exp. Medicine), 126, 423-4.
42 (1976)]. Class 4 and 6 cyclosporins inhibit the haemolysis zone compared to untreated controls at a concentration of 0.03 to 10.0 μg / ml. 4.2 Lymphocyte stimulation test by Janossey and Greaves [“Clin. Exp. Immunor”
unol.), 9, 483 (1971) and 10, 525.
P. (1972)]-Class 4 and 6 cyclosporins inhibit concanavalin A-stimulated DNA synthesis in mouse spleen lymphocytes compared to untreated controls at concentrations of 0.001-3.0 μg / ml (H ³ -Block thymidine incorporation), block cell proliferation and blastogenis.

4.3 Mixed lymphocyte reaction [Bach et al., "Journal of Experimental Medicine" (J. Ex.)
p. Med.), 136, 1430 (1972)]-atypical spleen cells (CBA) obtained from irradiated mice for 5 days.
-Incubated lymphocytes [mouse (valve
(Balb) / c) spleen cells] (ie, proliferation and differentiation) are measured in the presence and absence of the test substance. The reaction in the absence of the test substance was used as a control and was 100%
And The response in the presence of the test substance is expressed as a percentage change compared to a 100% control response. 0.001-3.0 μg / m
The inhibition of the reaction is observed with a concentration of l of class 4 and 6 cyclosporin.

Example 5 Anti-inflammatory activity Anti-inflammatory activity can be demonstrated in an adjuvant arthritis test in rats. For this test, the method of Pearson and Wood ["Arthr. Rheum."
2, 440 (1959)] to induce adjuvant arthritis. In this test, class 4 and 6 cyclosporins show activity against progressive and chronic arthritis at doses of 5-30 mg / kg / day (orally).

Example 6 Antiparasitic (antimalarial) activity lane, "Chemotherapy and Drug Resistance in Malaria"
ance in Malaria) (Peters ed., Academic Press, New York, 1970). On day 0, Plasmodiu Bergei
mberghei) (NK65 strain) with 0.2 ml (intraperitoneal administration) of a suspension containing 10 ⁷ parasite cells.
Male). On day 3, the test substance is administered subcutaneously at various doses using 5-10 mice / dose. The survival time is recorded and the minimum effective dose (MED) is calculated by comparing the survival time with that of the untreated control. For controls,
The survival time is about 7 days. MED is the dose that doubles the survival time. In this test, class 4 and 6 cyclosporins are effective at doses of 10-100 mg / kg / day.

Example 7 Effect on EAE in rats. 1. Effects on chronic EAE. Borel and others, "Agents and Actions" (A
According to the technique described by Gents and Actions, Vol. 6, page 468 (1976), EAE is induced in a group of 8-12 otherwise healthy, non-healthy male rats weighing 150-200 g. Rats are kept under laboratory conditions and have unlimited access to food and water. After 10 to 13 days, the onset of EAE is observed, for example, in the hind paws, paralytic symptoms are clearly observed. Test compounds are administered to test animals daily at a dose of 25-50 mg / kg (oral) for 5 consecutive days after the start of EAE. Rats are examined daily for signs of illness and the number of rats that have recovered and the day recovery has been observed. The observation is continued for another 5 to 8 weeks after the start of the treatment, and any recurrent cases are detected. The number of rats that recurred and the date of recurrence are recorded.

Following administration of class 4 and 6 cyclosporine in the above dose range, the reduction in recovery time compared to the control group receiving only placebo (olive oil) is recorded. 7.2 Action in preventing the occurrence of EAE. Perform the test in the same manner as described in 4.1 above. However, in this case, starting on the day of sensitization (inducing EAE),
The test compound is administered at a dose of 25-50 mg / kg (oral) daily for 4 days. Observe rats daily for signs of paralysis,
The start date of EAE in each affected rat is recorded.
EAs that may continue to be observed for several months
Detect late onset of E.

Following administration of Class 4 and 6 "cyclosporin" in the above dose range, the inhibition of the onset of EAE is observed during the observation period in comparison to controls treated with placebo (olive oil) alone. Class 4 and 6 cyclosporins are useful in the prevention and treatment of diseases and conditions that require a reduced immune response in view of their immunosuppressive activity. That is, by using them, for example, treatment of autoimmune disease or organ transplantation (for example, heart,
In preventing cardiopulmonary, skin, cornea, bone marrow, pancreas and kidney rejection, the proliferation of lymphocytes and immune cells can be suppressed.

Also, in view of their anti-inflammatory and EAE-related activities, class 4 and 6 cyclosporins are useful in the treatment of inflammatory conditions, especially inflammatory conditions with an etiology involving autoimmune components, such as arthritis (eg rheumatoid arthritis). , Chronic or progressive arthritis and osteoarthritis) and rheumatic diseases and autoimmune hematological disorders (including, for example, hemolytic anemia, anaplastic anemia, pure erythrocyte anemia and idiopathic thrombocytopenia), systemic lupus erythematosus,
Polychondritis, scleroderma, Wegner granulomatosis, dermatomyositis, chronic active hepatitis, myasthenia gravis, psoriasis, steven
-Johnson syndrome, idiopathic sprue, autoimmune inflammatory bowel disease (including ulcerative colitis and Crohn's disease), endocrine eye disease, Graves' disease, sarcoidosis, primary pericholangitis cirrhosis, primary juvenile diabetes
(Type I diabetes mellitus), uveitis (anterior and posterior), interstitial pulmonary fibrosis, psoriatic arthritis, glomerulonephritis (with and without nephrotic syndrome, e.g., idiopathic nephrotic syndrome or minimal changes Nephropathy) as well as in the treatment of multiple sclerosis.

Class 4 and 6 cyclosporins are also useful in treating parasitic infections in view of their antiparasitic activity. For example, coccidioidomycosis and schistosomiasis, and especially protozoal infections, especially malaria. Of course, for the above uses the dosage will depend on the cyclosporin chosen, the mode of administration, the particular condition to be treated and the therapy desired. However, in general, about 1-10
0, preferably about 5-50, most preferably 10-20
Administration at an oral daily dose of mg / kg (animal body weight) gives satisfactory results. In the case of large mammals (eg, humans), the total daily dose will be from about 75 to about 2000, preferably about 20
0 to about 1000, most preferably about 300 to about 700 mg
Within the range. Dosage forms suitable for oral administration may comprise from about 20 to about 20 to 50 mixed with a solid or liquid pharmaceutical diluent or carrier.
It contains about 2000, preferably about 50 to about 1000, most preferably about 75 to 700 mg of cyclosporine, and is conveniently administered in one or two to four divided doses daily.

As already indicated, a suitable daily dose rate is:
In particular, it depends on the particular cyclosporin chosen, for example, taking into account the relative potency of the action. For the preferred cyclosporins of classes 4 and 6, the results [IC ₅₀ values (μg / ml)] obtained in the series of experiments according to Example 4 above are as follows:

[Table 9] A suitable cyclosporin for use in antiparasitic indications is cyclosporin 4.7. The ED ₅₀ value obtained for this cyclosporin in the method of Example 6 is about 15.
0 mg / kg.

As described above, the present invention also provides the following. 7. A pharmaceutical composition comprising the above class 4 or 6 cyclosporin together with a pharmaceutically acceptable diluent or carrier. 8. A method of inducing immunosuppression in the treatment of an inflammatory disease or condition or the treatment of a parasitic infection in a subject in need thereof, comprising administering to said subject an effective amount of said class 4 or 6 cyclosporin. And 9. Class 4 or 6 cyclosporin used as a medicament, for example as an immunosuppressant, anti-inflammatory or antiparasitic agent.

Particular immunosuppressive, anti-inflammatory and anti-parasitic uses to which class 4 and 6 cyclosporins may be applied include, for example, the specific forms of organ transplantation, inflammatory diseases, autoimmune diseases or parasitic infections listed above. Including all of the foregoing uses.

──────────────────────────────────────────────────続 き Continued on the front page (51) Int.Cl. ⁶ Identification code FI A61K 38/00 AEB A61K 37/02 ADZ C07K 7/64 ZNA AEB (31) Priority claim number 8714098 (32) Priority date 1987 1987 March 17 (33) Priority claim country United Kingdom (GB) (31) Priority claim number 8714115 (32) Priority date June 17, 1987 (33) Priority claim country United Kingdom (GB) (31) Priority claim Number 8714118 (32) Priority date June 17, 1987 (33) Priority country United Kingdom (GB) (31) Priority claim number 8714119 (32) Priority date June 17, 1987 (33) Priority country United Kingdom (GB) (31) Priority claim number 8714125 (32) Priority date June 17, 1987 (33) Priority claiming country United Kingdom (GB) (72) Inventor Manfred Kriegel Zeha-4422 Alicedorf, Hauptstrasse 91 (72) Inventor Trebol Grin Peine Zeha-3005 Bern, Dharma Chilein 26 (72) Inventor René Pe Travel Zeha-4056 Basel, Switzerland No. 11, Bill Helm-His-Strasse (72) Inventor Laurent Wenger, No. 45, Grenzacherberg, Zeha-4125 Lichen, Switzerland (56) References JP-A-56-128725 (JP, A) JP-A-Showa 61-212599 (JP, A) (58) Fields investigated (Int. Cl. ⁶ , DB name) A61K 38/13 C07K 7/64 CA (STN)

Claims (1)

(57) [Claims] 1. A compound of the formula (II) Wherein A is a compound of the formula (VII) B represents -αAbu-, -Thr-, -Val-, -Nva
-Or β-O-acyl-α-amino acid residue, X is -Sar-, Y is -Val-], wherein the resistance to chemotherapeutic treatment is eliminated or reduced, or An agent that restores sensitivity to chemotherapy treatment.